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Affinity proteomics reveals extensive phosphorylation of the Brassica chromosome axis protein ASY 1 and a network of associated proteins at prophase I of meiosis
Author(s) -
Osman Kim,
Yang Jianhua,
Roitinger Elisabeth,
Lambing Christophe,
Heckmann Stefan,
Howell Elaine,
Cuacos Maria,
Imre Richard,
Dürnberger Gerhard,
Mechtler Karl,
Armstrong Susan,
Franklin F. Christopher H.
Publication year - 2018
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.13752
Subject(s) - biology , meiocyte , meiosis , proteomics , genetics , proteome , prophase , synaptonemal complex , chromosome , gene , microbiology and biotechnology , computational biology
Summary During meiosis, the formation of crossovers ( CO s) generates genetic variation and provides physical links that are essential for accurate chromosome segregation. CO s occur in the context of a proteinaceous chromosome axis. The transcriptomes and proteomes of anthers and meiocytes comprise several thousand genes and proteins, but because of the level of complexity relatively few have been functionally characterized. Our understanding of the physical and functional interactions between meiotic proteins is also limited. Here we use affinity proteomics to analyse the proteins that are associated with the meiotic chromosome axis protein, ASY 1, in Brassica oleracea anthers and meiocytes. We show that during prophase I ASY 1 and its interacting partner, ASY 3, are extensively phosphorylated, and we precisely assign phosphorylation sites. We identify 589 proteins that co‐immunoprecipitate with ASY 1. These correspond to 492 Arabidopsis orthologues, over 90% of which form a coherent protein–protein interaction ( PPI ) network containing known and candidate meiotic proteins, including proteins more usually associated with other cellular processes such as DNA replication and proteolysis. Mutant analysis confirms that affinity proteomics is a viable strategy for revealing previously unknown meiotic proteins, and we show how the PPI network can be used to prioritise candidates for analysis. Finally, we identify another axis‐associated protein with a role in meiotic recombination. Data are available via ProteomeXchange with identifier PXD 006042.