A complex pattern of H2A phosphorylation in the mouse testis

• Published in: Experimental cell research Vol. 195; no. 1; pp. 8 - 12
• Main Authors: Green, G.R., Patel, J.C., Hecht, N.B., Poccia, D.L.
• Format: Journal Article
• Language: English
• Published: Orlando, FL Elsevier Inc 01.07.1991; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Electrophoresis, Gel, Two-Dimensional; Fundamental and applied biological sciences. Psychology; Histones - chemistry; Histones - metabolism; Male; Mice; Peptide Mapping; Phosphoproteins - chemistry; Phosphoproteins - metabolism; Phosphorylation; Phosphoserine - metabolism; Proteins; Testis - metabolism; Vertebrata; Phosphorylation; Mammalia; Histone H2A; Mouse; Rodentia; Tissue culture; Metabolism; Testicle; Nuclear protein
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/0014-4827(91)90493-E

Phosphorylation of H2A histones in mouse testis was examined using testis tubule cultures labeled with 32PO 4. Histones were analyzed by two systems of two-dimensional polyacrylamide gel electrophoresis, followed by autoradiography of the gels. Of the 32PO 4 detected in histones, 95% was incorporated by certain modified forms of the H2A variants H2A.1 and H2A.X. Phosphorylation sites were mapped to N- and C-terminal regions of the modified variants by SDS gel electrophoresis and autoradiography of peptides generated by cleavage of in vitro-labeled proteins with N-bromosuccinimide. Incorporation rates differed for N- and C-terminal regions from different modified forms, demonstrating a complex pattern of H2A phosphorylation in the mouse testis.