Photochemical crosslinking of transcription complexes with psoralen. I. Covalent attachment of in vitro SV4O nascent RNA to its double-stranded DNA template

Che Kun James Shen, John E. Hearst

研究成果: 雜誌貢獻文章同行評審

7 引文 斯高帕斯(Scopus)

摘要

14c-labeled SV40 DNA has been transcribed with E. coli RNA polymerase using 3H-labeled ribonucleotide triphosphates as precursors. The resulting transcription complexes were then photochemically crosslinked with the psoralen derivative, 4′ -aminomethyl-4,5′ ,8-trimethylpsoralen (AMT), at 37°C and analyzed in SDS-sucrose gradients. It was found that the photochemical crosslinking procedure caused the nascent RNA chains to co-sediment with their double-stranded (helical) Sv40 templates in the denaturing sucrose gradient. This result and several control experiments suggest that covalent linkages have formed between nascent RNA and helical DNA after the photochemical reaction. The crosslinking phenomenon was observed to be independent of the superhelical state of the DNA used as the template. Prior addition of EDTA to stop the transcription is not requiréd for successful crosslinkage.

原文英語
頁(從 - 到)1429-1442
頁數14
期刊Nucleic Acids Research
5
發行號4
DOIs
出版狀態已發佈 - 四月 1978
對外發佈Yes

ASJC Scopus subject areas

  • Genetics

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