ABA-regulated SNRK2 Protein Kinase - Revision history

Alice Harmon at 16:21, 26 July 2014

Alice Harmon — Sat, 26 Jul 2014 16:21:57 GMT

←Older revision		Revision as of 16:21, 26 July 2014
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	\|'''Right scene''' - SnRK2.6 (blue) in complex with the protein phosphatase 2C, HAB1 (gold), with Mg<sup>2+</sup> and SO<sub>4</sub><sup>2-</sup> [[3ujg]]<ref name = "Soon2012"/>		\|'''Right scene''' - SnRK2.6 (blue) in complex with the protein phosphatase 2C, HAB1 (gold), with Mg<sup>2+</sup> and SO<sub>4</sub><sup>2-</sup> [[3ujg]]<ref name = "Soon2012"/>
	\|-		\|-
-	\| <applet load='3ujg' size='400' frame='true' align='left' caption='[[3uc4]] - Apo SnRK2.6' scene = '55/559985/Aposnrk2_6/6' /><br clear='both'>'''3uc4 scenes''' <Br><scene name='55/559985/Aposnrk2_6/7'>1. Default scene</scene> The catalytic domain of SnRK2.6 is typical of [[Eukaryotic Protein Kinase Catalytic Domain]]) except for an additional α-helix (shown as strands) in the small lobe, which is formed by SNRK2 box sequence. <Br><scene name='55/559985/Aposnrk2_6critical/3'>2. Important structures:</scene> The activation segment (with unresolved gap), including the D of the DFG motif in ball and stick, is blue. The catalytic loop, including the D of the DLKLEN motif in ball and stick, is orchid. Subdomain III, including its invariant E in ball and stick, is gold. The invariant K of subdomain II is in chartreuse. The SnRK2 box is turquoise. The C-terminal domain, that includes the ABA box is unresolved. The arrangement of the residues in ball and stick around the active site, indicate that this structure is in a partially active state in spite of its unphosphorylated activation loop. This is possibly due to the interaction of the SNRK2 box helix with subdomain III.<ref name = "Ng2011"/>. The interaction between these helices is similar to the interaction of helices in the complex between <scene name='55/559985/Cdk2-cyclin/3'>cyclin-dependent protein kinase 2 (CDK2) and cyclin</scene> [[1w98]]. Here we see that subdomain III of the protein kinase (opaque blue) is stablized by interaction with a helix from cyclin (opaque gold). The positioning of subdomain III by this interaction is critical for for formation of the active site.<ref>PMID:15660127</ref>. <br><br><br><br><br><br><br><br><br><br>	+	\| <applet name= 'one' load='3ujg' size='400' frame='true' align='left' caption='[[3uc4]] - Apo SnRK2.6' scene = '55/559985/Aposnrk2_6/6' /><br clear='both'>'''3uc4 scenes''' <Br><scene name='55/559985/Aposnrk2_6/7' target= 'one'>1. Default scene</scene><scene name='55/559985/Aposnrk2_6/7' target= 'one'>1. Default scene</scene> The catalytic domain of SnRK2.6 is typical of [[Eukaryotic Protein Kinase Catalytic Domain]]) except for an additional α-helix (shown as strands) in the small lobe, which is formed by SNRK2 box sequence. <Br><scene name='55/559985/Aposnrk2_6critical/3' target= 'one'>2. Important structures:</scene> The activation segment (with unresolved gap), including the D of the DFG motif in ball and stick, is blue. The catalytic loop, including the D of the DLKLEN motif in ball and stick, is orchid. Subdomain III, including its invariant E in ball and stick, is gold. The invariant K of subdomain II is in chartreuse. The SnRK2 box is turquoise. The C-terminal domain, that includes the ABA box is unresolved. The arrangement of the residues in ball and stick around the active site, indicate that this structure is in a partially active state in spite of its unphosphorylated activation loop. This is possibly due to the interaction of the SNRK2 box helix with subdomain III.<ref name = "Ng2011"/>. The interaction between these helices is similar to the interaction of helices in the complex between <scene name='55/559985/Cdk2-cyclin/3' target= 'one'>cyclin-dependent protein kinase 2 (CDK2) and cyclin</scene> [[1w98]]. Here we see that subdomain III of the protein kinase (opaque blue) is stablized by interaction with a helix from cyclin (opaque gold). The positioning of subdomain III by this interaction is critical for for formation of the active site.<ref>PMID:15660127</ref>. <br><br><br><br><br><br><br><br><br><br>
-	\| <applet load='3ujg' size='400' frame='true' align='left' caption='[[3ujg]] - SnRK2.6-HAB1' scene = '55/559985/Aposnrk2_6/2' /><Br clear='both'>'''3ujg scenes'''<Br><scene name='55/559985/Aposnrk2_6/2'>1. Default Scene</scene> The two enzymes are bound via interface their active sites. The phosphatase inactivates the kinase by dephosphorylating the kinase activation loop and by sterically blocking the kinase active site. The complex was constructed as a fusion protein with a 6His-tag at the N-terminus of SnRK2.6 (residues 11–362) fused to HAB1(172–511) via a GSGSAGSAAGS linker. Mutations of D296A and E297A in SnRK2.6 were introduced at the crystal packing interface to reduce surface entropy. <br><scene name='55/559985/Ost1hab1_critical/3'>2. Important structures in SnRK2.6</scene> The same structures as in the left scene are shown. The fully resolved activation segment extends into the phosphatase's active site and is unphosphorylated. Residues 319-362 of SnRK2.6, which includes the ABA box, and the GSGSAGSAAGS linker are not resolved. The disorganization of the residues shown in ball and stick, with most pointing away from the active site, indicates that the catalytic domain is in the inactive state.<Br><scene name='55/559985/Ost1hab1_interaction/2'>3. Zone of interaction</scene> The activation loop (blue trace) of SnRK2.6 is inserted into the catalytic site (marked by the magnesium ions) of the phosphatase. The phosphorylatable residue of the activation loop S175 (CPK ball and stick) is positioned near the magnesium ions. W385 of the phosphatase (brown ball and stick) in turn protrudes into the kinase's active site, where it interacts with residues R139 and Glu144 (CPK ball and stick) of the catalytic loop (orchid trace) and I183 of the activation loop.<br><scene name='55/559985/Tetherbinding/1'> 5. Proposed interaction zone </scene> SnRK2.6 is shown in blue cartoon, and HAB1 in gold spacefill. The ABA box sequence is not resolved, but it would extend from the C-terminal end of the SNRK2 box helix (cyan helix). It is proposed that the ABA box sequence, which is highly acidic, binds to a patch of basic residues (blue) on the surface of the phosphatase.<ref name= "Soon2012"/>	+	\| <applet name= 'two' load='3ujg' size='400' frame='true' align='left' caption='[[3ujg]] - SnRK2.6-HAB1' scene = '55/559985/Aposnrk2_6/2' /><Br clear='both'>'''3ujg scenes'''<Br><scene name='55/559985/Aposnrk2_6/2' target= 'two'>1. Default Scene</scene><scene name='55/559985/Aposnrk2_6/2' target= 'two'>1. Default Scene</scene> The two enzymes are bound via interface their active sites. The phosphatase inactivates the kinase by dephosphorylating the kinase activation loop and by sterically blocking the kinase active site. The complex was constructed as a fusion protein with a 6His-tag at the N-terminus of SnRK2.6 (residues 11–362) fused to HAB1(172–511) via a GSGSAGSAAGS linker. Mutations of D296A and E297A in SnRK2.6 were introduced at the crystal packing interface to reduce surface entropy. <br><scene name='55/559985/Ost1hab1_critical/3' target= 'two'>2. Important structures in SnRK2.6</scene> The same structures as in the left scene are shown. The fully resolved activation segment extends into the phosphatase's active site and is unphosphorylated. Residues 319-362 of SnRK2.6, which includes the ABA box, and the GSGSAGSAAGS linker are not resolved. The disorganization of the residues shown in ball and stick, with most pointing away from the active site, indicates that the catalytic domain is in the inactive state.<Br><scene name='55/559985/Ost1hab1_interaction/2' target= 'two'>3. Zone of interaction</scene> The activation loop (blue trace) of SnRK2.6 is inserted into the catalytic site (marked by the magnesium ions) of the phosphatase. The phosphorylatable residue of the activation loop S175 (CPK ball and stick) is positioned near the magnesium ions. W385 of the phosphatase (brown ball and stick) in turn protrudes into the kinase's active site, where it interacts with residues R139 and Glu144 (CPK ball and stick) of the catalytic loop (orchid trace) and I183 of the activation loop.<br><scene name='55/559985/Tetherbinding/1' target= 'two'> 4. Proposed interaction zone </scene> SnRK2.6 is shown in blue cartoon, and HAB1 in gold spacefill. The ABA box sequence is not resolved, but it would extend from the C-terminal end of the SNRK2 box helix (cyan helix). It is proposed that the ABA box sequence, which is highly acidic, binds to a patch of basic residues (blue) on the surface of the phosphatase.<ref name= "Soon2012"/>
	\|}		\|}

Michal Harel at 11:51, 5 January 2014

Michal Harel — Sun, 05 Jan 2014 11:51:22 GMT

←Older revision		Revision as of 11:51, 5 January 2014
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	\|'''Right scene''' - SnRK2.6 (blue) in complex with the protein phosphatase 2C, HAB1 (gold), with Mg<sup>2+</sup> and SO<sub>4</sub><sup>2-</sup> [[3ujg]]<ref name = "Soon2012"/>		\|'''Right scene''' - SnRK2.6 (blue) in complex with the protein phosphatase 2C, HAB1 (gold), with Mg<sup>2+</sup> and SO<sub>4</sub><sup>2-</sup> [[3ujg]]<ref name = "Soon2012"/>
	\|-		\|-
-	\| <applet load='3ujg' size='400' frame='true' align='left' caption='3uc4 - Apo SnRK2.6' scene = '55/559985/Aposnrk2_6/6' /><br clear='both'>'''3uc4 scenes''' <Br><scene name='55/559985/Aposnrk2_6/7'>1. Default scene</scene> The catalytic domain of SnRK2.6 is typical of [[Eukaryotic Protein Kinase Catalytic Domain]]) except for an additional α-helix (shown as strands) in the small lobe, which is formed by SNRK2 box sequence. <Br><scene name='55/559985/Aposnrk2_6critical/3'>2. Important structures:</scene> The activation segment (with unresolved gap), including the D of the DFG motif in ball and stick, is blue. The catalytic loop, including the D of the DLKLEN motif in ball and stick, is orchid. Subdomain III, including its invariant E in ball and stick, is gold. The invariant K of subdomain II is in chartreuse. The SnRK2 box is turquoise. The C-terminal domain, that includes the ABA box is unresolved. The arrangement of the residues in ball and stick around the active site, indicate that this structure is in a partially active state in spite of its unphosphorylated activation loop. This is possibly due to the interaction of the SNRK2 box helix with subdomain III.<ref name = "Ng2011"/>. The interaction between these helices is similar to the interaction of helices in the complex between <scene name='55/559985/Cdk2-cyclin/3'>cyclin-dependent protein kinase 2 (CDK2) and cyclin</scene> [[1w98]]. Here we see that subdomain III of the protein kinase (opaque blue) is stablized by interaction with a helix from cyclin (opaque gold). The positioning of subdomain III by this interaction is critical for for formation of the active site.<ref>PMID:15660127</ref>. <br><br><br><br><br><br><br><br><br><br>	+	\| <applet load='3ujg' size='400' frame='true' align='left' caption='[[3uc4]] - Apo SnRK2.6' scene = '55/559985/Aposnrk2_6/6' /><br clear='both'>'''3uc4 scenes''' <Br><scene name='55/559985/Aposnrk2_6/7'>1. Default scene</scene> The catalytic domain of SnRK2.6 is typical of [[Eukaryotic Protein Kinase Catalytic Domain]]) except for an additional α-helix (shown as strands) in the small lobe, which is formed by SNRK2 box sequence. <Br><scene name='55/559985/Aposnrk2_6critical/3'>2. Important structures:</scene> The activation segment (with unresolved gap), including the D of the DFG motif in ball and stick, is blue. The catalytic loop, including the D of the DLKLEN motif in ball and stick, is orchid. Subdomain III, including its invariant E in ball and stick, is gold. The invariant K of subdomain II is in chartreuse. The SnRK2 box is turquoise. The C-terminal domain, that includes the ABA box is unresolved. The arrangement of the residues in ball and stick around the active site, indicate that this structure is in a partially active state in spite of its unphosphorylated activation loop. This is possibly due to the interaction of the SNRK2 box helix with subdomain III.<ref name = "Ng2011"/>. The interaction between these helices is similar to the interaction of helices in the complex between <scene name='55/559985/Cdk2-cyclin/3'>cyclin-dependent protein kinase 2 (CDK2) and cyclin</scene> [[1w98]]. Here we see that subdomain III of the protein kinase (opaque blue) is stablized by interaction with a helix from cyclin (opaque gold). The positioning of subdomain III by this interaction is critical for for formation of the active site.<ref>PMID:15660127</ref>. <br><br><br><br><br><br><br><br><br><br>
-	\| <applet load='3ujg' size='400' frame='true' align='left' caption='3ujg - SnRK2.6-HAB1' scene = '55/559985/Aposnrk2_6/2' /><Br clear='both'>'''3ujg scenes'''<Br><scene name='55/559985/Aposnrk2_6/2'>1. Default Scene</scene> The two enzymes are bound via interface their active sites. The phosphatase inactivates the kinase by dephosphorylating the kinase activation loop and by sterically blocking the kinase active site. The complex was constructed as a fusion protein with a 6His-tag at the N-terminus of SnRK2.6 (residues 11–362) fused to HAB1(172–511) via a GSGSAGSAAGS linker. Mutations of D296A and E297A in SnRK2.6 were introduced at the crystal packing interface to reduce surface entropy. <br><scene name='55/559985/Ost1hab1_critical/3'>2. Important structures in SnRK2.6</scene> The same structures as in the left scene are shown. The fully resolved activation segment extends into the phosphatase's active site and is unphosphorylated. Residues 319-362 of SnRK2.6, which includes the ABA box, and the GSGSAGSAAGS linker are not resolved. The disorganization of the residues shown in ball and stick, with most pointing away from the active site, indicates that the catalytic domain is in the inactive state.<Br><scene name='55/559985/Ost1hab1_interaction/2'>3. Zone of interaction</scene> The activation loop (blue trace) of SnRK2.6 is inserted into the catalytic site (marked by the magnesium ions) of the phosphatase. The phosphorylatable residue of the activation loop S175 (CPK ball and stick) is positioned near the magnesium ions. W385 of the phosphatase (brown ball and stick) in turn protrudes into the kinase's active site, where it interacts with residues R139 and Glu144 (CPK ball and stick) of the catalytic loop (orchid trace) and I183 of the activation loop.<br><scene name='55/559985/Tetherbinding/1'> 5. Proposed interaction zone </scene> SnRK2.6 is shown in blue cartoon, and HAB1 in gold spacefill. The ABA box sequence is not resolved, but it would extend from the C-terminal end of the SNRK2 box helix (cyan helix). It is proposed that the ABA box sequence, which is highly acidic, binds to a patch of basic residues (blue) on the surface of the phosphatase.<ref name= "Soon2012"/>	+	\| <applet load='3ujg' size='400' frame='true' align='left' caption='[[3ujg]] - SnRK2.6-HAB1' scene = '55/559985/Aposnrk2_6/2' /><Br clear='both'>'''3ujg scenes'''<Br><scene name='55/559985/Aposnrk2_6/2'>1. Default Scene</scene> The two enzymes are bound via interface their active sites. The phosphatase inactivates the kinase by dephosphorylating the kinase activation loop and by sterically blocking the kinase active site. The complex was constructed as a fusion protein with a 6His-tag at the N-terminus of SnRK2.6 (residues 11–362) fused to HAB1(172–511) via a GSGSAGSAAGS linker. Mutations of D296A and E297A in SnRK2.6 were introduced at the crystal packing interface to reduce surface entropy. <br><scene name='55/559985/Ost1hab1_critical/3'>2. Important structures in SnRK2.6</scene> The same structures as in the left scene are shown. The fully resolved activation segment extends into the phosphatase's active site and is unphosphorylated. Residues 319-362 of SnRK2.6, which includes the ABA box, and the GSGSAGSAAGS linker are not resolved. The disorganization of the residues shown in ball and stick, with most pointing away from the active site, indicates that the catalytic domain is in the inactive state.<Br><scene name='55/559985/Ost1hab1_interaction/2'>3. Zone of interaction</scene> The activation loop (blue trace) of SnRK2.6 is inserted into the catalytic site (marked by the magnesium ions) of the phosphatase. The phosphorylatable residue of the activation loop S175 (CPK ball and stick) is positioned near the magnesium ions. W385 of the phosphatase (brown ball and stick) in turn protrudes into the kinase's active site, where it interacts with residues R139 and Glu144 (CPK ball and stick) of the catalytic loop (orchid trace) and I183 of the activation loop.<br><scene name='55/559985/Tetherbinding/1'> 5. Proposed interaction zone </scene> SnRK2.6 is shown in blue cartoon, and HAB1 in gold spacefill. The ABA box sequence is not resolved, but it would extend from the C-terminal end of the SNRK2 box helix (cyan helix). It is proposed that the ABA box sequence, which is highly acidic, binds to a patch of basic residues (blue) on the surface of the phosphatase.<ref name= "Soon2012"/>
	\|}		\|}

Alice Harmon at 17:54, 5 October 2013

Alice Harmon — Sat, 05 Oct 2013 17:54:20 GMT

←Older revision		Revision as of 17:54, 5 October 2013
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	Three members of the SnRK2 family of protein kinases - SnRK2.6/OST1/SRK2E, SNRK2.2/SRK2D and SnRK2.3/SRK2I - are activated by the [[ABA Signaling Pathway]]<ref name ="Umezawa2009"> PMID:19805022 </ref><ref name ="Soon2012"> PMID:22116026 </ref><ref name = "Mustilli2002">PMID:12468729</ref><ref>PMID:12514244</ref><ref name = "Nakashima2009">PMID:19541597</ref><ref name = "Fujii2007">PMID:17307925</ref> . SnRK2 stands for <u>Sn</u>f1-<u>r</u>elated protein <u>k</u>inase family, group <u>2</u>. These protein kinases have [[Eukaryotic Protein Kinase Catalytic Domain]]s that are related to yeast Snf1, and they belong to the calmodulin-dependent protein kinase family of the kinome.		Three members of the SnRK2 family of protein kinases - SnRK2.6/OST1/SRK2E, SNRK2.2/SRK2D and SnRK2.3/SRK2I - are activated by the [[ABA Signaling Pathway]]<ref name ="Umezawa2009"> PMID:19805022 </ref><ref name ="Soon2012"> PMID:22116026 </ref><ref name = "Mustilli2002">PMID:12468729</ref><ref>PMID:12514244</ref><ref name = "Nakashima2009">PMID:19541597</ref><ref name = "Fujii2007">PMID:17307925</ref> . SnRK2 stands for <u>Sn</u>f1-<u>r</u>elated protein <u>k</u>inase family, group <u>2</u>. These protein kinases have [[Eukaryotic Protein Kinase Catalytic Domain]]s that are related to yeast Snf1, and they belong to the calmodulin-dependent protein kinase family of the kinome.

-	~~Two of~~ SnRK2.6/OST1/SRK2E's three names originated from its membership in subclass III of the SnRK2 family of protein kinases. It was named SnRK2.6 by Hrabak et al.<ref>PMID:12805596</ref> and SRK2E by Umezawa et al.<ref name ="Umezawa2009"/>. The third name OST1 (open stomata 1)<ref name = "Mustilli2002"/> is descriptive of the phenotype of plants bearing a gene mutation that produces an inactive protein kinase.	+	The best studied ABA-regulated protein kinase is SnRK2.6/OST1/SRK2E. Two of its three names originated from its membership in subclass III of the SnRK2 family of protein kinases. It was named SnRK2.6 by Hrabak et al.<ref>PMID:12805596</ref> and SRK2E by Umezawa et al.<ref name ="Umezawa2009"/>. The third name OST1 (open stomata 1)<ref name = "Mustilli2002"/> is descriptive of the phenotype of plants bearing a gene mutation that produces an inactive protein kinase.

	In rice homologs of these protein kinases are named SAPK8, SAPK9 and SAPK10.		In rice homologs of these protein kinases are named SAPK8, SAPK9 and SAPK10.

-	==Kinase ~~structure and~~ regulation==	+	==Kinase regulation and structure==
		+
		+	As shown in [[ABA Signaling Pathway]] SnRK2.6/OST1/SRK2E and its homologs are regulated indirectly by ABA. Kinase activity is doubly inhibited in the absence of ABA by its binding to an [[ABA-regulated Protein Phosphatase 2C]]. This interaction results in dephosphorylation of the protein kinase's activation loop and blocking of its active site by phosphatase. When ABA binds to its receptor, the receptor binds to the protein phosphatase, freeing the protein kinase. The kinase is now free to be activated by phosphorylation of its activation loop by either itself or another protein kinase.

	SnRK2.6/OST1/SRK2E has a primary structure comprising an amino terminal [[Eukaryotic Protein Kinase Catalytic Domain]] and a C-terminal sequence that contains the SNRK2 box, which is unique to the SNRK2 family and required for activity<ref name= "Ng2011"/><ref name ="Belin2006"> PMID: 16766677</ref>. Its C-terminus also contains a sequence called the ABA box, which is found only in the family members that are responsive to ABA<ref name ="Belin2006">. The latter sequence is required for binding to PP2C<ref name ="Belin2006">, but is not seen in the crystal structure.		SnRK2.6/OST1/SRK2E has a primary structure comprising an amino terminal [[Eukaryotic Protein Kinase Catalytic Domain]] and a C-terminal sequence that contains the SNRK2 box, which is unique to the SNRK2 family and required for activity<ref name= "Ng2011"/><ref name ="Belin2006"> PMID: 16766677</ref>. Its C-terminus also contains a sequence called the ABA box, which is found only in the family members that are responsive to ABA<ref name ="Belin2006">. The latter sequence is required for binding to PP2C<ref name ="Belin2006">, but is not seen in the crystal structure.
		+
		+	The scenes below explore the structure of SnRK2.6/OST1/SRK2E as a free monomer and in complex with protein phosphatase 2C, HAB1

	<ref name ="Soon2013"> PMID:22116026 </ref>		<ref name ="Soon2013"> PMID:22116026 </ref>

Alice Harmon at 17:36, 5 October 2013

Alice Harmon — Sat, 05 Oct 2013 17:36:53 GMT

←Older revision		Revision as of 17:36, 5 October 2013
Line 1:		Line 1:
	==Kinase names and family members==		==Kinase names and family members==
-	Three members of the SnRK2 family of protein kinases - SnRK2.6/OST1/SRK2E, SNRK2.2/SRK2D and SnRK2.3/SRK2I - are activated by the [[ABA Signaling Pathway]]. SnRK2 stands for <u>Sn</u>f1-<u>r</u>elated protein <u>k</u>inase family, group <u>2</u>. These protein kinases have [[Eukaryotic Protein Kinase Catalytic Domain]]s that are related to yeast Snf1, and they belong to the calmodulin-dependent protein kinase family of the kinome.	+	Three members of the SnRK2 family of protein kinases - SnRK2.6/OST1/SRK2E, SNRK2.2/SRK2D and SnRK2.3/SRK2I - are activated by the [[ABA Signaling Pathway]]<ref name ="Umezawa2009"> PMID:19805022 </ref><ref name ="Soon2012"> PMID:22116026 </ref><ref name = "Mustilli2002">PMID:12468729</ref><ref>PMID:12514244</ref><ref name = "Nakashima2009">PMID:19541597</ref><ref name = "Fujii2007">PMID:17307925</ref> . SnRK2 stands for <u>Sn</u>f1-<u>r</u>elated protein <u>k</u>inase family, group <u>2</u>. These protein kinases have [[Eukaryotic Protein Kinase Catalytic Domain]]s that are related to yeast Snf1, and they belong to the calmodulin-dependent protein kinase family of the kinome.

	Two of SnRK2.6/OST1/SRK2E's three names originated from its membership in subclass III of the SnRK2 family of protein kinases. It was named SnRK2.6 by Hrabak et al.<ref>PMID:12805596</ref> and SRK2E by Umezawa et al.<ref name ="Umezawa2009"/>. The third name OST1 (open stomata 1)<ref name = "Mustilli2002"/> is descriptive of the phenotype of plants bearing a gene mutation that produces an inactive protein kinase.		Two of SnRK2.6/OST1/SRK2E's three names originated from its membership in subclass III of the SnRK2 family of protein kinases. It was named SnRK2.6 by Hrabak et al.<ref>PMID:12805596</ref> and SRK2E by Umezawa et al.<ref name ="Umezawa2009"/>. The third name OST1 (open stomata 1)<ref name = "Mustilli2002"/> is descriptive of the phenotype of plants bearing a gene mutation that produces an inactive protein kinase.

Alice Harmon at 17:28, 5 October 2013

Alice Harmon — Sat, 05 Oct 2013 17:28:14 GMT

←Older revision		Revision as of 17:28, 5 October 2013
Line 1:		Line 1:
	==Kinase names and family members==		==Kinase names and family members==
-	~~Two~~ of SnRK2.6/OST1/SRK2E~~'s three names originated from its membership in subclass III of the SnRK2 family of protein kinases~~. ~~It was named~~ SnRK2.6 by ~~Hrabak et al~~.<~~ref~~>~~PMID:12805596~~</~~ref~~> ~~and SRK2E by Umezawa et al.~~<~~ref name ="Umezawa2009"~~/>~~. SnRK2 stands for SNF1-related kinase~~ group 2~~, which in Arabidopsis has 10 members~~. ~~SNRK2s~~ are ~~members of~~ the calmodulin-dependent protein kinase ~~clade of protein kinases. The third name OST1 (open stomata 1)<ref name = "Mustilli2002"/> is descriptive~~ of the ~~phenotype of plants bearing a gene mutation that produces an inactive protein kinase~~.	+	Three members of the SnRK2 family of protein kinases - SnRK2.6/OST1/SRK2E, SNRK2.2/SRK2D and SnRK2.3/SRK2I - are activated by the [[ABA Signaling Pathway]]. SnRK2 stands for <u>Sn</u>f1-<u>r</u>elated protein <u>k</u>inase family, group <u>2</u>. These protein kinases have [[Eukaryotic Protein Kinase Catalytic Domain]]s that are related to yeast Snf1, and they belong to the calmodulin-dependent protein kinase family of the kinome.

-	Two ~~other family members in Arabidopsis, SNRK2.2/SRK2D and~~ SnRK2.3/~~SRK2I, are activated by the ABA pathway~~ in the ~~same manner as~~ SnRK2.6. ~~Each of these kinases interacts with a member of clade A~~ of the protein ~~phosphatase 2C family - ABI1, HAB1 or HAB2~~. In rice homologs of these protein kinases are named SAPK8, SAPK9 and SAPK10.	+	Two of SnRK2.6/OST1/SRK2E's three names originated from its membership in subclass III of the SnRK2 family of protein kinases. It was named SnRK2.6 by Hrabak et al.<ref>PMID:12805596</ref> and SRK2E by Umezawa et al.<ref name ="Umezawa2009"/>. The third name OST1 (open stomata 1)<ref name = "Mustilli2002"/> is descriptive of the phenotype of plants bearing a gene mutation that produces an inactive protein kinase.
		+
		+	In rice homologs of these protein kinases are named SAPK8, SAPK9 and SAPK10.

	==Kinase structure and regulation==		==Kinase structure and regulation==

Alice Harmon: New page: ==Kinase names and family members== Two of SnRK2.6/OST1/SRK2E's three names originated from its membership in subclass III of the SnRK2 family of protein kinases. It was named SnRK2.6 by H...

Alice Harmon — Sat, 05 Oct 2013 17:12:47 GMT

New page: ==Kinase names and family members== Two of SnRK2.6/OST1/SRK2E's three names originated from its membership in subclass III of the SnRK2 family of protein kinases. It was named SnRK2.6 by H...

New page

==Kinase names and family members==
Two of SnRK2.6/OST1/SRK2E's three names originated from its membership in subclass III of the SnRK2 family of protein kinases. It was named SnRK2.6 by Hrabak et al.<ref>PMID:12805596</ref> and SRK2E by Umezawa et al.<ref name ="Umezawa2009"/>. SnRK2 stands for SNF1-related kinase group 2, which in Arabidopsis has 10 members. SNRK2s are members of the calmodulin-dependent protein kinase clade of protein kinases. The third name OST1 (open stomata 1)<ref name = "Mustilli2002"/> is descriptive of the phenotype of plants bearing a gene mutation that produces an inactive protein kinase.

Two other family members in Arabidopsis, SNRK2.2/SRK2D and SnRK2.3/SRK2I, are activated by the ABA pathway in the same manner as SnRK2.6. Each of these kinases interacts with a member of clade A of the protein phosphatase 2C family - ABI1, HAB1 or HAB2. In rice homologs of these protein kinases are named SAPK8, SAPK9 and SAPK10.

==Kinase structure and regulation==

SnRK2.6/OST1/SRK2E has a primary structure comprising an amino terminal [[Eukaryotic Protein Kinase Catalytic Domain]] and a C-terminal sequence that contains the SNRK2 box, which is unique to the SNRK2 family and required for activity<ref name= "Ng2011"/><ref name ="Belin2006"> PMID: 16766677</ref>. Its C-terminus also contains a sequence called the ABA box, which is found only in the family members that are responsive to ABA<ref name ="Belin2006">. The latter sequence is required for binding to PP2C<ref name ="Belin2006">, but is not seen in the crystal structure.

<ref name ="Soon2013"> PMID:22116026 </ref>

{|
|'''Left scene''' - unphosphorylated SnRK2.6 without any ligands [[3uc4]]<ref name = "Ng2011">PMID:22160701</ref>
|'''Right scene''' - SnRK2.6 (blue) in complex with the protein phosphatase 2C, HAB1 (gold), with Mg2+ and SO42- [[3ujg]]<ref name = "Soon2012"/>
|-
| <applet load='3ujg' size='400' frame='true' align='left' caption='3uc4 - Apo SnRK2.6' scene = '55/559985/Aposnrk2_6/6' /> '''3uc4 scenes''' <scene name='55/559985/Aposnrk2_6/7'>1. Default scene</scene> The catalytic domain of SnRK2.6 is typical of [[Eukaryotic Protein Kinase Catalytic Domain]]) except for an additional α-helix (shown as strands) in the small lobe, which is formed by SNRK2 box sequence. <scene name='55/559985/Aposnrk2_6critical/3'>2. Important structures:</scene> The activation segment (with unresolved gap), including the D of the DFG motif in ball and stick, is blue. The catalytic loop, including the D of the DLKLEN motif in ball and stick, is orchid. Subdomain III, including its invariant E in ball and stick, is gold. The invariant K of subdomain II is in chartreuse. The SnRK2 box is turquoise. The C-terminal domain, that includes the ABA box is unresolved. The arrangement of the residues in ball and stick around the active site, indicate that this structure is in a partially active state in spite of its unphosphorylated activation loop. This is possibly due to the interaction of the SNRK2 box helix with subdomain III.<ref name = "Ng2011"/>. The interaction between these helices is similar to the interaction of helices in the complex between <scene name='55/559985/Cdk2-cyclin/3'>cyclin-dependent protein kinase 2 (CDK2) and cyclin</scene> [[1w98]]. Here we see that subdomain III of the protein kinase (opaque blue) is stablized by interaction with a helix from cyclin (opaque gold). The positioning of subdomain III by this interaction is critical for for formation of the active site.<ref>PMID:15660127</ref>. 
| <applet load='3ujg' size='400' frame='true' align='left' caption='3ujg - SnRK2.6-HAB1' scene = '55/559985/Aposnrk2_6/2' /> '''3ujg scenes''' <scene name='55/559985/Aposnrk2_6/2'>1. Default Scene</scene> The two enzymes are bound via interface their active sites. The phosphatase inactivates the kinase by dephosphorylating the kinase activation loop and by sterically blocking the kinase active site. The complex was constructed as a fusion protein with a 6His-tag at the N-terminus of SnRK2.6 (residues 11–362) fused to HAB1(172–511) via a GSGSAGSAAGS linker. Mutations of D296A and E297A in SnRK2.6 were introduced at the crystal packing interface to reduce surface entropy. <scene name='55/559985/Ost1hab1_critical/3'>2. Important structures in SnRK2.6</scene> The same structures as in the left scene are shown. The fully resolved activation segment extends into the phosphatase's active site and is unphosphorylated. Residues 319-362 of SnRK2.6, which includes the ABA box, and the GSGSAGSAAGS linker are not resolved. The disorganization of the residues shown in ball and stick, with most pointing away from the active site, indicates that the catalytic domain is in the inactive state. <scene name='55/559985/Ost1hab1_interaction/2'>3. Zone of interaction</scene> The activation loop (blue trace) of SnRK2.6 is inserted into the catalytic site (marked by the magnesium ions) of the phosphatase. The phosphorylatable residue of the activation loop S175 (CPK ball and stick) is positioned near the magnesium ions. W385 of the phosphatase (brown ball and stick) in turn protrudes into the kinase's active site, where it interacts with residues R139 and Glu144 (CPK ball and stick) of the catalytic loop (orchid trace) and I183 of the activation loop. <scene name='55/559985/Tetherbinding/1'> 5. Proposed interaction zone </scene> SnRK2.6 is shown in blue cartoon, and HAB1 in gold spacefill. The ABA box sequence is not resolved, but it would extend from the C-terminal end of the SNRK2 box helix (cyan helix). It is proposed that the ABA box sequence, which is highly acidic, binds to a patch of basic residues (blue) on the surface of the phosphatase.<ref name= "Soon2012"/>
|}

===SNRK2 structures===

[[3uc3]] ''Arabidopsis thaliana'' SNRK2.3 + Co2+ 
[[3zut]] AtSNRK2.6 (D160A mutant)+ ANP 
[[3zuu]] AtSNRK2.6 (D160A, S175D mutant) + gold 
[[3uc4]] apoAtSNRK2.6 (D59A, E60A mutant) 
[[3udb]] apoAtSNRK2.6 (C131A, C157A, C159A, S7A, s29A, s43A, S166A, T175A) 

'''complex with a protein phosphatase 2C''' 
[[3ujg]] AtSNRK2.6 (D296A) + HAB1 + Mg2+

==References==

<references/>

==See Also==
[http://en.wikipedia.org/wiki/Abscisic_acid] Abscisic Acid in Wikipedia