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Involvement of C-terminal structural elements of Equine Infectious Anemia Virus reverse transcriptase in DNA polymerase and ribonuclease H activities

Title data

Rausch, Jason W. ; Arts, Eric J. ; Wöhrl, Birgitta M. ; Le Grice, Stuart F.J.:
Involvement of C-terminal structural elements of Equine Infectious Anemia Virus reverse transcriptase in DNA polymerase and ribonuclease H activities.
In: Journal of Molecular Biology. Vol. 257 (1996) Issue 3 . - pp. 500-511.
ISSN 0022-2836
DOI: https://doi.org/10.1006/jmbi.1996.0181

Abstract in another language

In order to investigate the modes of DNA synthesis supported by the 66 and 51 kDa subunits of equine infectious anemia virus reverse transcriptase (EIAV RT), recombinant p66 polypeptides containing a modified ribonuclease H (RNase H) domain were purified and evaluated. Defined heteropolymeric template-primer combinations and high-resolution gel electrophoresis provided a qualitative evaluation of DNA polymerase and RNase H activities, while DNase I footprinting revealed features of replication complexes containing the truncated enzymes. Removal of α-helix E′ and the conserved β5′-αE′ “His-loop” in p66Δ20 RT uncouples the RNase H activities, alters affinity for template-primer and dictates how the replicating enzyme responds to secondary structure on both DNA and RNA templates. Despite these alterations, DNase I footprinting shows no major difference in the overall structure of DNA-directed DNA synthesis complexes. In contrast, removing 47 C-terminal residues, which includes α-helix D′, β-strand 5′ and α-Helix E′, yields an enzyme with distributive DNA polymerase properties closely resembling the purified p51 subunit.

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 Biopolymers
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biopolymers > Lehrstuhl Biopolymere - Apl. Prof. Dr. Birgitta Wöhrl
Result of work at the UBT: No
DDC Subjects: 500 Science > 540 Chemistry
500 Science > 570 Life sciences, biology
Date Deposited: 20 May 2019 07:30
Last Modified: 20 May 2019 07:30
URI: https://eref.uni-bayreuth.de/id/eprint/48997