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  • The Initiation of Meiotic Sex Chromosome Inactivation Sequesters DNA Damage Signaling from Autosomes in Mouse Spermatogenesis.

The Initiation of Meiotic Sex Chromosome Inactivation Sequesters DNA Damage Signaling from Autosomes in Mouse Spermatogenesis.

Current biology : CB (2020-01-07)
Hironori Abe, Kris G Alavattam, Yueh-Chiang Hu, Qishen Pang, Paul R Andreassen, Rashmi S Hegde, Satoshi H Namekawa
ABSTRACT

Meiotic sex chromosome inactivation (MSCI) is an essential event in the mammalian male germline. MSCI is directed by a DNA damage response (DDR) pathway centered on the phosphorylation of histone variant H2AX at serine 139 (termed γH2AX). The failure to initiate MSCI is linked to complete meiotic arrest and elimination of germ cells; however, the mechanisms underlying this arrest and elimination remain unknown. To address this question, we established a new separation-of-function mouse model for H2ax that shows specific and complete defects in MSCI. The genetic change is a point mutation in which another H2AX amino acid residue important in the DDR, tyrosine 142 (Y142), is converted to alanine (H2ax-Y142A). In H2ax-Y142A meiosis, the establishment of DDR signals on the chromosome-wide domain of the sex chromosomes is impaired. The initiation of MSCI is required for stage progression, which enables crossover formation, suggesting that the establishment of MSCI permits the timely progression of male meiosis. Our results suggest that normal meiotic progression requires the removal of ATR-mediated DDR signaling from autosomes. We propose a novel biological function for MSCI: the initiation of MSCI sequesters DDR factors from autosomes to the sex chromosomes at the onset of the pachytene stage, and the subsequent formation of an isolated XY nuclear compartment-the XY body-sequesters DDR factors to permit meiotic progression from the mid-pachytene stage onward. VIDEO ABSTRACT.

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Millipore
Membrana Immobilon®-P in PVDF, 1 roll, 27 cm x 3.75 m, 0.45 µm pore size, Hydrophobic PVDF Transfer Membrane for western blotting.
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Cocktail di inibitori delle proteasi 2, aqueous solution (dark coloration may develop upon storage, which does not affect the activity)
Roche
Kit per la rilevazione della morte cellulare in situ, fluoresceina, sufficient for ≤50 tests, suitable for detection
Sigma-Aldrich
Anticorpo anti fosfo-istone H2A.X (Ser139), colone JBW301, clone JBW301, Upstate®, from mouse
Sigma-Aldrich
Anti-Rad51 (Ab-1) Rabbit pAb, liquid, Calbiochem®
Supelco
Cloruro di calcio, 0.1 M Ca, analytical standard (for ion-selective electrodes)
Sigma-Aldrich
Anti-phospho Histone H2A.X (Ser139) Antibody, clone JBW301, Alexa Fluor 647, clone JBW301, 0.5 mg/mL, from mouse
Millipore
ReBlot Plus Kit