Proteomics

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LC-MSMS of Arabidopsis thaliana flower bud samples (TurboID-based proximity labeling of meiotic chromosome axes)


ABSTRACT: The meiotic chromosome axis is a proteinaceous structure along which sister chromatids are arranged in a loop-base array during meiotic prophase I. In diverse species including plants, chromosome axis components have been shown to be critical for synapsis and meiotic recombination and hence essential for assuring the fidelity of meiosis. In Arabidopsis thaliana, the limited number of meiotic cells embedded in floral organs constrains proteomic approaches for the identification of new axis components. Therefore, we employed TurboID (TbID) based proximity labeling (PL) in meiotic cells of A. thaliana. The two axis proteins ASY1 and ASY3 were selected and ASY1-TbID and ASY3-TbID transgenic plants were generated to enable specific targeted biotin labeling of meiotic chromosome axis. ASY1-eYFP and nucTbID (TbID driven by UBQ10 promoter) were generated and used as controls. These lines were either treated or not with exogenous biotin followed by streptavidin affinity-purification coupled with mass spectrometry to identify their proximate candidate proteins. By stringent filtering a final candidates list was established which covered most of the known meiotic chromosome axis-related proteins and new proteins without a reported meiotic function. Functional validation of selected candidates and comparison with ASY1 AP-MS data (in Brassica oleracea) suggest the TbID based PL is a robust tool for identification of proximate proteins in meiotic cells of A. thaliana.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Flower Bud

SUBMITTER: Richard Imre  

LAB HEAD: Stefan Heckmann

PROVIDER: PXD034241 | Pride | 2023-03-08

REPOSITORIES: Pride

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