Role of T7 RNA polymerase His784 in start site selection and initial transcription

Luis G. Brieba, Robert Padilla, Rui J Sousa

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The role of steric constraints vs sequence preference in start site selection by T7 RNA polymerase was investigated by using a series of synthetic promoters in which the preferred template strand 'CC' initiation sequence was moved away from its normal position relative to the -17 to -6 element of the T7 promoter. It was found that the CC sequence directs efficient initiation if placed 1 or 2 nt downstream of its normal position, but not if placed upstream, or more than 2 nt downstream, of +1. Mutagenesis revealed that part of the bias to initiate with GTP is due to an interaction between histidine 784 and the 2-amino group of a guanosine bound in the initiating triphosphate position. This interaction is also important for holding short transcripts within the transcription complex during initial transcription.

Original languageEnglish (US)
Pages (from-to)5144-5149
Number of pages6
JournalBiochemistry
Volume41
Issue number16
DOIs
StatePublished - Apr 23 2002

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Site selection
Guanosine
Transcription
Guanosine Triphosphate
Histidine
Mutagenesis
bacteriophage T7 RNA polymerase
triphosphoric acid

ASJC Scopus subject areas

  • Biochemistry

Cite this

Role of T7 RNA polymerase His784 in start site selection and initial transcription. / Brieba, Luis G.; Padilla, Robert; Sousa, Rui J.

In: Biochemistry, Vol. 41, No. 16, 23.04.2002, p. 5144-5149.

Research output: Contribution to journalArticle

Brieba, Luis G. ; Padilla, Robert ; Sousa, Rui J. / Role of T7 RNA polymerase His784 in start site selection and initial transcription. In: Biochemistry. 2002 ; Vol. 41, No. 16. pp. 5144-5149.
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