5ekf
From Proteopedia
Crystallization and X-ray Diffraction Data Collection of Importin-alpha from Mus musculus Complexed with a XPG NLS Peptide, fragment 1
Structural highlights
DiseaseERCC5_HUMAN Xeroderma pigmentosum complementation group G;COFS syndrome. The disease is caused by mutations affecting the gene represented in this entry. FunctionERCC5_HUMAN Single-stranded structure-specific DNA endonuclease involved in DNA excision repair. Makes the 3'incision in DNA nucleotide excision repair (NER). Acts as a cofactor for a DNA glycosylase that removes oxidized pyrimidines from DNA. May also be involved in transcription-coupled repair of this kind of damage, in transcription by RNA polymerase II, and perhaps in other processes too. Publication Abstract from PubMedXeroderma pigmentosum type G (XPG) proteins are involved in DNA lesion recognition and promotion of nucleotide excision repair (NER). Specific mutations in these proteins may lead to Cockayne syndrome, in which the patients may display severe developmental retardation and neurological abnormalities. No structural information is available for their spacer region or the C-terminal domain, which are important, respectively, for specific NER activity and substrate specificity, and nuclear translocation. Immunofluorescence studies suggested two specific regions of the XPG C-terminus as potential bipartite nuclear localization sequences, which would be responsible for its translocation to the nucleus by the classical nuclear import pathway mediated by importin-alpha (Impalpha). Thus, in order to test these hypotheses and gain insight into the structural basis for the nuclear import process for the XPG protein, we solved the crystal structures of complexes formed by the Impalpha and peptides corresponding to both putative NLS (nuclear localization signal) sequences (XPG1 and XPG2) and performed isothermal titration calorimetry (ITC) assays to determine their binding affinities. Structural experiments confirm the binding of both NLS peptides to Impalpha but, unexpectedly, they bind to the receptor as monopartite NLSs. The ITC assays demonstrated that XPG1 and XPG2 peptides bind to two separate binding-sites, but with high affinity to the major NLS binding-site of the Impalpha, resembling classical monopartite SV40 TAg NLS. The results lead to insights about what distinguishes monopartite and bipartite NLSs, as well as the differential roles of XPG1 and XPG2 NLSs in the nuclear localization of XPG. Structural and calorimetric studies demonstrate that Xeroderma pigmentosum type G (XPG) can be imported to the nucleus by a classical nuclear import pathway via a monopartite NLS sequence.,de Barros AC, Takeda AA, Dreyer TR, Velazquez-Campoy A, Kobe B, Fontes MR J Mol Biol. 2016 Jan 23. pii: S0022-2836(16)00038-3. doi:, 10.1016/j.jmb.2016.01.019. PMID:26812207[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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