Abstract:
:A temperature-sensitive mutant of Escherichia coli affected in the alpha subunit of RNA polymerase has been investigated. Gene mapping and complementation experiments placed the mutation to temperature-sensitivity within the alpha operon at 72 min. on the bacterial chromosome. The rate of RNA synthesis in vivo and the accumulation of ribosomal RNA were significantly reduced in the mutant at 44 degrees C. The thermostability at 44 degrees C of the purified holoenzyme from mutant cells was about 20% of that of the normal enzyme. Assays with T7 DNA as a template showed that the fraction of active enzyme competent for transcription was reduced as a function of assay temperature but that initiation and elongation were not significantly affected by the alpha mutation. A major effect on the fidelity of transcription was observed with the mutant enzyme, with misincorporation on two different templates stimulated about 4 fold at 37 degrees C. The role of the alpha dimer in the structure and function of RNA polymerase is discussed.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Mehrpouyan M,Champney WSdoi
10.1093/nar/18.12.3445subject
Has Abstractpub_date
1990-06-25 00:00:00pages
3445-50issue
12eissn
0305-1048issn
1362-4962journal_volume
18pub_type
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