Preparation of efficient excision repair competent cell-free extracts from C. reinhardtii cells

Chaudhari, Vishalsingh ; Raghavan, Vandana ; Rao, Basuthkar J. (2014) Preparation of efficient excision repair competent cell-free extracts from C. reinhardtii cells PLoS One, 9 (10). Article ID e109160, 7 pages. ISSN 1932-6203

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Official URL: http://journals.plos.org/plosone/article?id=10.137...

Related URL: http://dx.doi.org/10.1371/journal.pone.0109160

Abstract

Chlamydomonas reinhardtii is a prospective model system for understanding molecular mechanisms associated with DNA repair in plants and algae. To explore this possibility, we have developed an in vitro repair system from C. reinhardtii cell-free extracts that can efficiently repair UVC damage (Thymine-dimers) in the DNA. We observed that excision repair (ER) synthesis based nucleotide incorporation, specifically in UVC damaged supercoiled (SC) DNA, was followed by ligation of nicks. Photoreactivation efficiently competed out the ER in the presence of light. In addition, repair efficiency in cell-free extracts from ER deficient strains was several fold lower than that of wild-type cell extract. Interestingly, the inhibitor profile of repair DNA polymerase involved in C. reinhardtii in vitro ER system was akin to animal rather than plant DNA polymerase. The methodology to prepare repair competent cell-free extracts described in the current study can aid further molecular characterization of ER pathway in C. reinhardtii.

Item Type:Article
Source:Copyright of this article belongs to Public Library of Science.
ID Code:106609
Deposited On:16 Jun 2017 11:02
Last Modified:16 Jun 2017 11:02

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