Open AccessIdentification of Histone Messenger RNA from HeLa Cells
Author(s) -
Breindl Michael,
GALLWTTZ Dieter
Publication year - 1973
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1973.tb02620.x
Subject(s) - microbiology and biotechnology , histone , rna , biology , messenger rna , polysome , reticulocyte , uridine , polyacrylamide gel electrophoresis , thymidine , dna , hela , biochemistry , in vitro , ribosome , gene , enzyme
Histone mRNA from HeLa cells synchronized by a double thymidine block has been identified and characterized. Its synthesis starts immediately after the cells are released from the thymidine block. Pulse labelling for 60 min with [5‐ 3 H]uridine at different times of the S‐phase showed that radioactively labelled histone mRNA enters polyribosomes with a maximum during the first hour of S‐phase, i.e. , before the maximum of DNA synthesis. Histone mRNA sediments on sucrose gradients in the 8–10‐S region and separates on polyacrylamide gels into three RNA species with molecular weights of about 1.5 × 10 5 , 1.8 × 10 6 , and 2.1 × 10 5 , respectively. Polyribosomal 8–10‐S RNA from synchronized cells in S‐phase directs the synthesis of histones in a rabbit reticulocyte cell‐free system. Histones synthesized in vitro were identified by polyacrylamide gel electrophoresis and by high‐voltage paper electrophoresis of tryptic peptides. Polyribosomal 8–10‐S RNA from cells in which DNA synthesis was blocked by thymidine or hydroxyurea had only a marginal histone mRNA activity.