EFFICIENT TRANSCRIPTION OF A DNA-TEMPLATE ASSOCIATED WITH HISTONE (H3-CENTER-DOT-H4)(2) TETRAMERS

被引:0
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作者
PUERTA, C [1 ]
HERNANDEZ, F [1 ]
GUTIERREZ, C [1 ]
PINEIRO, M [1 ]
LOPEZALARCON, L [1 ]
PALACIAN, E [1 ]
机构
[1] UNIV AUTONOMA MADRID, CSIC, CTR BIOL MOLEC SEVERO OCHOA, E-28049 MADRID, SPAIN
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中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A histone-DNA transcription template has been assembled, by dialysis against decreasing salt concentrations, from pGEMEX-1 (4 kilobases), a plasmid containing a promoter for bacteriophage T7 RNA polymerase, and from isolated histone (H3.H4)2 tetramers. Electron microscopy after psoralen cross-linking shows that each histone tetramer protects approximately 80 base pairs of DNA from psoralen action and that, under the employed conditions, an average of 15 tetramer particles are assembled per DNA molecule. This (H3.H4)2-DNA template is efficiently transcribed in vitro by T7 RNA polymerase as compared to naked DNA. The presence of (H3-H4)2 tetramers does not affect initiation, in contrast with the complete histone octamer, (H2A.H2B.H3.H4)2, assembled with the complementary addition of H2A-H2B dimers, which causes transcriptional inhibition mainly by blocking initiation.
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页码:26663 / 26667
页数:5
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