RRC ID 87690
著者 Chelysheva L, Grandont L, Vrielynck N, le Guin S, Mercier R, Grelon M.
タイトル An easy protocol for studying chromatin and recombination protein dynamics during Arabidopsis thaliana meiosis: immunodetection of cohesins, histones and MLH1.
ジャーナル Cytogenet Genome Res
Abstract Plant meiosis studies have enjoyed a fantastic boom in recent years with the use of Arabidopsis thaliana as a model not only for molecular genetics and genomics but also for cytogenetics. In this article we describe a new protocol for immunolabelling meiotic proteins that allows the detection of a large range of proteins on strongly spread chromosomes throughout the entire meiotic process. We used this method to immunodetect MLH1, a crucial component of the meiotic recombination machinery, and found that it can be visualised as foci from pachytene to diakinesis, where it co-localises with chiasmata. The mean MLH1 foci number per meiotic cell at diakinesis was 8.4 for WS-4 and 9.95 for Col-0, with the number of foci per bivalent ranging from 1 to 5. We also analysed MLH1 distribution within bivalents and found that they were not restricted to specific chromosomal regions. The analysis of MLH1 foci formation in the Atzip4 mutant, where class I crossover (CO) formation is prevented, revealed that residual chiasmata were not labelled by MLH1, strongly suggesting that MLH1 antibodies only label class I COs in Arabidopsis. It thus appears that the 'obligatory CO' is systematically labeled by MLH1 and is generated through the class I pathway.
巻・号 129(1-3)
ページ 143-53
公開日 2010-7-1
DOI 10.1159/000314096
PII 000314096
PMID 20628250
MeSH Adenosine Triphosphatases / immunology Adenosine Triphosphatases / metabolism Animals Antibodies Arabidopsis / cytology* Arabidopsis / genetics* Arabidopsis / metabolism Arabidopsis Proteins / immunology Arabidopsis Proteins / metabolism* Cell Cycle Proteins / metabolism Chromatin / genetics Chromatin / metabolism Chromosomal Proteins, Non-Histone / metabolism Chromosomes, Plant / genetics Chromosomes, Plant / metabolism Cohesins Crossing Over, Genetic Cytogenetic Analysis / methods* Histones / metabolism Immunohistochemistry / methods* Meiosis / genetics* Meiosis / physiology MutL Protein Homolog 1 Rats Recombination, Genetic
IF 1.114
リソース情報
シロイヌナズナ / 植物培養細胞・遺伝子 pda02005