| RRC ID |
87690
|
| Author |
Chelysheva L, Grandont L, Vrielynck N, le Guin S, Mercier R, Grelon M.
|
| Title |
An easy protocol for studying chromatin and recombination protein dynamics during Arabidopsis thaliana meiosis: immunodetection of cohesins, histones and MLH1.
|
| Journal |
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.
|
| Volume |
129(1-3)
|
| Pages |
143-53
|
| Published |
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
|
| Resource |
| Arabidopsis / Cultured plant cells, genes |
pda02005 |
