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Mutating the "primer grip" of p66 HIV-1 reverse transcriptase implicates Tryptophan-229 in template-primer utilization

Title data

Jacques, Pamela S. ; Wöhrl, Birgitta M. ; Ottmann, Michele ; Darlix, Jean-Luc ; Le Grice, Stuart F.J.:
Mutating the "primer grip" of p66 HIV-1 reverse transcriptase implicates Tryptophan-229 in template-primer utilization.
In: The Journal of Biological Chemistry. Vol. 269 (1994) Issue 42 . - pp. 26472-26478.
ISSN 1083-351X

Official URL: Volltext

Abstract in another language

"BcgI cassette" mutagenesis was used to prepare variants of p66 human immunodeficiency virus (HIV)-1 reverse transcriptase with amino acid substitutions between residues Glu224 and Trp229. Mutant polypeptides were reconstituted in vitro with wild type p51 to generate the "selectively mutated" heterodimer series p66(224A)/p51-p66(229A)/p51. Purified enzymes were characterized with respect to dimerization, DNA polymerase, RNase H, and tRNA(Lys-3) binding. The combined analyses indicate that while alteration of p66 residues Glu224-Leu228 has minimal consequences, the DNA polymerase activities of mutant p66(229A)/p51 are impaired. DNase I footprinting illustrates that this mutant does not form a stable replication complex with a model template-primer. In vivo studies indicate that the equivalent mutation eliminates viral infectivity, suggesting a contribution of Trp229 toward architecture of the p66 primer grip.

Further data

Item Type: Article in a journal
Refereed: Yes
Institutions of the University: Faculties > Faculty of Biology, Chemistry and Earth Sciences
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry with an Emphasis on Biophysical Chemistry
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Former Professors > Chair Biopolymers - Apl. Prof. Dr. Birgitta Wöhrl
Faculties
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Former Professors
Result of work at the UBT: No
DDC Subjects: 500 Science > 540 Chemistry
500 Science > 570 Life sciences, biology
Date Deposited: 20 May 2019 11:48
Last Modified: 20 May 2019 11:48
URI: https://eref.uni-bayreuth.de/id/eprint/49004