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4w7s

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Revision as of 22:11, 4 August 2016

Crystal structure of the yeast DEAD-box splicing factor Prp28 at 2.54 Angstroms resolution

Structural highlights

4w7s is a 2 chain structure. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:	, , ,
NonStd Res:
Activity:	RNA helicase, with EC number 3.6.4.13
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

[PRP28_YEAST] ATP-dependent RNA helicase involved in mRNA splicing. May destabilize the U1/5'-splice site duplex to permit an effective competition for the 5'-splice site by the U6 snRNA, resulting in the switch between U1 and U6 at the 5'-splice site. May also act to unwind the U4/U6 base-pairing interaction in the U4/U6/U5 snRNP, facilitating the first covalent step of splicing.^[1] ^[2] ^[3]

Publication Abstract from PubMed

Yeast Prp28 is a DEAD-box pre-mRNA splicing factor implicated in displacing U1 snRNP from the 5' splice site. Here we report that the 588-aa Prp28 protein consists of a trypsin-sensitive 126-aa N-terminal segment (of which aa 1-89 are dispensable for Prp28 function in vivo) fused to a trypsin-resistant C-terminal catalytic domain. Purified recombinant Prp28 and Prp28-(127-588) have an intrinsic RNA-dependent ATPase activity, albeit with a low turnover number. The crystal structure of Prp28-(127-588) comprises two RecA-like domains splayed widely apart. AMPPNP*Mg2+ is engaged by the proximal domain, with proper and specific contacts from Phe194 and Gln201 (Q motif) to the adenine nucleobase. The triphosphate moiety of AMPPNP*Mg2+ is not poised for catalysis in the open domain conformation. Guided by the Prp28*AMPPNP structure, and that of the Drosophila Vasa*AMPPNP*Mg2+*RNA complex, we targeted 20 positions in Prp28 for alanine scanning. ATP-site components Asp341 and Glu342 (motif II) and Arg527 and Arg530 (motif VI) and RNA-site constituent Arg476 (motif Va) are essential for Prp28 activity in vivo. Synthetic lethality of double-alanine mutations highlighted functionally redundant contacts in the ATP-binding (Phe194-Gln201, Gln201-Asp502) and RNA-binding (Arg264-Arg320) sites. Overexpression of defective ATP-site mutants, but not defective RNA-site mutants, elicited severe dominant-negative growth defects.

Crystal structure, mutational analysis and RNA-dependent ATPase activity of the yeast DEAD-box pre-mRNA splicing factor Prp28.,Jacewicz A, Schwer B, Smith P, Shuman S Nucleic Acids Res. 2014 Oct 10. pii: gku930. PMID:25303995^[4]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Staley JP, Guthrie C. An RNA switch at the 5' splice site requires ATP and the DEAD box protein Prp28p. Mol Cell. 1999 Jan;3(1):55-64. PMID:10024879
↑ Chen JY, Stands L, Staley JP, Jackups RR Jr, Latus LJ, Chang TH. Specific alterations of U1-C protein or U1 small nuclear RNA can eliminate the requirement of Prp28p, an essential DEAD box splicing factor. Mol Cell. 2001 Jan;7(1):227-32. PMID:11172727
↑ Strauss EJ, Guthrie C. A cold-sensitive mRNA splicing mutant is a member of the RNA helicase gene family. Genes Dev. 1991 Apr;5(4):629-41. PMID:2010088
↑ Jacewicz A, Schwer B, Smith P, Shuman S. Crystal structure, mutational analysis and RNA-dependent ATPase activity of the yeast DEAD-box pre-mRNA splicing factor Prp28. Nucleic Acids Res. 2014 Oct 10. pii: gku930. PMID:25303995 doi:http://dx.doi.org/10.1093/nar/gku930

1 Structural highlights
2 Function
3 Publication Abstract from PubMed
4 See Also
5 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/4w7s"

@@ Line 1: / Line 1: @@
 ==Crystal structure of the yeast DEAD-box splicing factor Prp28 at 2.54 Angstroms resolution==
 <StructureSection load='4w7s' size='340' side='right' caption='[[4w7s]], [[Resolution|resolution]] 2.54&Aring;' scene=''>
@@ Line 6: / Line 7: @@
 <tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=MSE:SELENOMETHIONINE'>MSE</scene></td></tr>
 <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/RNA_helicase RNA helicase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.6.4.13 3.6.4.13] </span></td></tr>
-<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4w7s FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4w7s OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4w7s RCSB], [http://www.ebi.ac.uk/pdbsum/4w7s PDBsum]</span></td></tr>
+<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4w7s FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4w7s OCA], [http://pdbe.org/4w7s PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4w7s RCSB], [http://www.ebi.ac.uk/pdbsum/4w7s PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4w7s ProSAT]</span></td></tr>
 </table>
 == Function ==
@@ Line 18: / Line 19: @@
 From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
 </div>
+<div class="pdbe-citations 4w7s" style="background-color:#fffaf0;"></div>
+==See Also==
+*[[Helicase|Helicase]]
 == References ==
 <references/>

4w7s

From Proteopedia

Revision as of 22:11, 4 August 2016

Crystal structure of the yeast DEAD-box splicing factor Prp28 at 2.54 Angstroms resolution

Structural highlights

Function

Publication Abstract from PubMed

See Also

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

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