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粟酒裂殖酵母蛋白提取物中两种错配结合活性的鉴定。

Identification of two mismatch-binding activities in protein extracts of Schizosaccharomyces pombe.

作者信息

Fleck O, Schär P, Kohli J

机构信息

Institute of General Microbiology, University of Bern, Switzerland.

出版信息

Nucleic Acids Res. 1994 Dec 11;22(24):5289-95. doi: 10.1093/nar/22.24.5289.

DOI:10.1093/nar/22.24.5289
PMID:7816618
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC332073/
Abstract

We have performed band-shift assays to identify mismatch-binding proteins in cell extracts of Schizosaccharomyces pombe. By testing heteroduplex DNA containing either a T/G or a C/C mismatch, two distinct band shifts were produced in the gels. A low mobility complex was observed with the T/G substrate, while a high mobility complex was present with C/C. Further analysis of the mismatch-binding specificities revealed that the T/G binding activity also binds to T/C, C/T, T/T, T/-, A/-, C/-, G/-, G/G, A/A, A/C, A/G, G/T, G/A, and C/A substrates with varying efficiencies, but not binds to C/C. The C/C binding activity efficiently binds to C/C, T/C, C/T, C/A, A/C, C/-, and weakly also to T/T, while all other mispairs are not recognized. Protein extracts of a mutant strain, defective in the mutS homologue swi4, displayed both mismatch-binding activities. Thus, swi4 does not encode for either one of the mismatch-binding proteins.

摘要

我们进行了凝胶迁移率变动分析，以鉴定粟酒裂殖酵母细胞提取物中的错配结合蛋白。通过检测含有T/G或C/C错配的异源双链DNA，在凝胶中产生了两种不同的条带迁移。用T/G底物观察到一个低迁移率复合物，而用C/C底物则存在一个高迁移率复合物。对错配结合特异性的进一步分析表明，T/G结合活性也以不同效率结合到T/C、C/T、T/T、T/-、A/-、C/-、G/-、G/G、A/A、A/C、A/G、G/T、G/A和C/A底物，但不结合C/C。C/C结合活性有效地结合C/C、T/C、C/T、C/A、A/C、C/-，也微弱地结合T/T，而所有其他错配均未被识别。在mutS同源物swi4中存在缺陷的突变菌株的蛋白质提取物显示出两种错配结合活性。因此，swi4不编码任何一种错配结合蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/74ee86ac75d4/nar00048-0155-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/34acfb7d2aec/nar00048-0153-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/e4d5fffbef82/nar00048-0153-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/cf3229d94de8/nar00048-0154-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/74ee86ac75d4/nar00048-0155-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/34acfb7d2aec/nar00048-0153-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/e4d5fffbef82/nar00048-0153-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/cf3229d94de8/nar00048-0154-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc9c/332073/74ee86ac75d4/nar00048-0155-a.jpg

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本文引用的文献

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