In vitro reconstitution of budding yeast pre-RC, competent in recruitment of recombinant pre-IC components, using purified, recombinant proteins

Kawakami, H., Stillman, B. (2012) In vitro reconstitution of budding yeast pre-RC, competent in recruitment of recombinant pre-IC components, using purified, recombinant proteins. Genes & Genetic Systems, 87 (6). p. 385. ISSN 1341-7568

URL: https://www.jstage.jst.go.jp/article/ggs/87/6/87_3...
DOI: 10.1266/ggs.87.375

Abstract

Pre-RC formation on chromosomal replication origins is an essential process for eukaryotic DNA replication in order for the origins to form pre-IC complexes. Though purification of each pre-RC component is of great importance to conduct quantitative analyses in vitro, recombinant MCM in budding yeast was a challenge to purify as a form active in pre-RC formation. We purified recombinant MCM complexed with Cdt1 and then reconstituted pre-RC formation using only purified, recombinant proteins. The established reaction could occur on representative early and late origins and was repressed by certain origin mutations. Reconstituted pre-RC had affinity for recombinant pre-IC components, indicating that a recombinant pre-IC could be assembled in vitro.

Item Type: Paper
Additional Information: Meeting Abstract
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > origin recognition complex
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
organism description > yeast
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: Matt Covey
Date Deposited: 25 Sep 2013 15:36
Last Modified: 20 Jun 2017 16:25
URI: http://repository.cshl.edu/id/eprint/28623

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