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Jeong W, Kang C. The histidine-805 in motif-C of the phage SP6 RNA polymerase is essential for its activity as revealed by random mutagenesis. Biochem Mol Biol Int. 1997;42(4):711-6doi: 10.1080/15216549700203131.
Jeong, W., & Kang, C. (1997). The histidine-805 in motif-C of the phage SP6 RNA polymerase is essential for its activity as revealed by random mutagenesis. Biochemistry and molecular biology international, 42(4), 711-6. https://doi.org/10.1080/15216549700203131
Jeong, W, and Kang, C. "The histidine-805 in motif-C of the phage SP6 RNA polymerase is essential for its activity as revealed by random mutagenesis." Biochemistry and molecular biology international vol. 42,4 (1997): 711-6. doi: https://doi.org/10.1080/15216549700203131
Jeong W, Kang C. The histidine-805 in motif-C of the phage SP6 RNA polymerase is essential for its activity as revealed by random mutagenesis. Biochem Mol Biol Int. 1997 Jul;42(4):711-6. doi: 10.1080/15216549700203131. PMID: 19856287.
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Biochem Mol Biol Int. 1997 Jul;42(4):711-6. doi: 10.1080/15216549700203131.

The histidine-805 in motif-C of the phage SP6 RNA polymerase is essential for its activity as revealed by random mutagenesis.

Biochemistry and molecular biology international

W Jeong, C Kang

Affiliations

  1. Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Taejon, Korea.

PMID: 19856287 DOI: 10.1080/15216549700203131

Abstract

In order to identify functional residues of the bacteriophage SP6 RNA polymerase, its C-terminal one-twelfth region was randomly mutagenized using polymerase chain reactions of its gene under the conditions for reduced fidelity of Taq DNA polymerase. Using a two-vector system that permits phenotypic isolation of mutants with reduced in vivo transcription activity, 3 single and 1 multiple mutants were isolated. A single substitution of Gln for His805 resulted in complete inactivation of the enzyme. A multiple mutant carrying substitutions at 808, 820, 835, 843 and 848 also abolished the activity. However, changes of Pro856-->Ser and Asp862-->Glu individually reduced the activity only slightly. It is noteworthy that His805 is one of the two motif-C residues that are absolutely conserved among all the DNA polymerases and monomeric RNA polymerases.

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