Dataset Information

Affinity proteomics targeting the meiotic chromosome axis protein, ASY1, from Brassica oleracea.

ABSTRACT: Meiosis is a highly complex process that underpins recombination in sexually reproducing organisms. Recent genomics studies suggest that up to several thousand genes/proteins contribute to the meiotic pathway, yet relatively few have been functionally characterised. Our understanding of the physical interactions between meiotic proteins and how the meiotic machinery interacts with other components of the cell is also limited. In this study, we used affinity proteomics targeting the meiotic chromosome axis protein, ASY1, to enrich for axis-associated proteins in Brassica oleracea anthers or meiocytes in prophase I of meiosis. LC-MS/MS analysis identified 540 proteins which co-immunoprecipitated with ASY1 in a sample-specific manner. These correspond to 485 Arabidopsis orthologues, 90% of which form a coherent predicted protein-protein interaction network which includes known and novel meiotic proteins, based on mutant analysis, but also proteins more usually associated with other cellular processes including replication and proteolysis. Our data provided new insights into the role of the chromosome axis in plant meiosis. We identified a novel axis-associated protein and showed that during prophase I, ASY1 and its interacting partner, ASY3, are extensively phosphorylated. Further studies targeting other meiotic proteins may ultimately enable the construction of a comprehensive meiosis protein-protein interaction network for higher plants.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Brassica Oleracea Var. Alboglabra

TISSUE(S): Anther, Plant Cell

SUBMITTER: Richard Imre

LAB HEAD: F. Chris H. Franklin

PROVIDER: PXD006042 | Pride | 2017-11-01

REPOSITORIES: Pride

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Publications

Affinity proteomics reveals extensive phosphorylation of the Brassica chromosome axis protein ASY1 and a network of associated proteins at prophase I of meiosis.

Osman Kim K Yang Jianhua J Roitinger Elisabeth E Lambing Christophe C Heckmann Stefan S Howell Elaine E Cuacos Maria M Imre Richard R Dürnberger Gerhard G Mechtler Karl K Armstrong Susan S Franklin F Christopher H FCH

The Plant journal : for cell and molecular biology 20171202 1

During meiosis, the formation of crossovers (COs) generates genetic variation and provides physical links that are essential for accurate chromosome segregation. COs 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 pro ...[more]

PMID: 29078019

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Project description:Dynamic nuclear architecture and chromatin organizations are the key features of the mid- prophase I in mammalian meiosis. Where the chromatin undergoes major changes, including meiosis-specific spatiotemporal arrangements and remodelling, the establishment of chromatin loop–axis structure, pairing, and crossing over between homologous chromosomes, any deficiencies in these events may induce genome instability, subsequently leading to failure to produce gametes and infertility. Despite the significance of chromatin structure, little is known about the location of chromatin marks and the necessity of their balance during meiosis prophase I. Here, we show a thorough cytological study of the surface-spread meiotic chromosomes of mouse spermatocytes for H3K9,14,18,23,27,36, H4K12,16 acetylation, and H3K4,9,27,36 methylation. Active acetylation and methylation marks on H3 and H4, such as H3K9ac, H3K14ac, H3K18ac, H3K36ac, H3K56ac, H4K12ac, H4K16ac, and H3K27me3 and H3K36me3 exhibited pan-nuclear localization away from heterochromatin. In comparison, repressive marks like H3K9me3 are localized to heterochromatin. In addition, we found that the intensities of H3K23ac, H3K27ac, and H3K9me3 enhanced during meiotic prophase I. Further, taking advantage of the delivery of small-molecule chemical inhibitors Methotrexate (MTX; heterochromatin enhancer), HMS-I1 (heterochromatin inhibitor), Anacardic acid (AA; histone acetyltransferase inhibitor), Trichostatin A (TSA; histone deacetylase inhibitor), IOX1 (JmjC demethylases inhibitor), and AZ505 (methyl transferase inhibitor) in seminiferous tubules through the rete testis route, revealed that alteration in histone modifications enhanced the centromere mislocalization, chromosome breakage, altered meiotic recombination and reduced sperm count. Notably, an imbalance in the histone modifications influences the chromosome axis length and chromatin loop size via transcriptional regulation of meiosis- specific genes. Our findings highlight the importance of balanced chromatin modifications in meiotic prophase I chromosome organization and instability.

Dataset Information

Affinity proteomics targeting the meiotic chromosome axis protein, ASY1, from Brassica oleracea.

Publications

Affinity proteomics reveals extensive phosphorylation of the Brassica chromosome axis protein ASY1 and a network of associated proteins at prophase I of meiosis.

Similar Datasets

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