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Processing of an HIV replication intermediate by the human DNA replication enzyme FEN1
J A Rumbaugh1, G M Fuentes, R A Bambara
1Department of Biochemistry, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
The Journal of Biological Chemistry
|October 24, 1998
Summary
Human flap endonuclease-1 (FEN1) cleaves overlaps during human immunodeficiency virus (HIV) replication. This enzyme
Area of Science:
- Molecular biology
- Virology
- Enzymology
Background:
- Human immunodeficiency virus (HIV) type 1 replication involves complex nucleic acid processing.
- Flap endonuclease-1 (FEN1) is a nuclease involved in DNA repair and replication.
Purpose of the Study:
- To investigate the role of human FEN1 in the replication of HIV type 1.
- To determine if FEN1 can process specific substrate structures arising during HIV replication.
Main Methods:
- Utilized model substrates mimicking HIV replication intermediates.
- Assessed the endonucleolytic cleavage activity of FEN1 on these substrates.
- Investigated the subsequent steps of DNA synthesis and ligation using DNA polymerase epsilon and DNA ligase I.
Main Results:
- FEN1 demonstrated endonucleolytic cleavage of primer-template substrates with a 5'-unannealed tail.
- FEN1 successfully removed the central (+)-strand overlap formed during HIV synthesis.
- Human DNA ligase I completed the double-strand synthesis after FEN1 processing and primer extension.
Conclusions:
- Human FEN1 plays a role in processing overlapping nucleic acid structures during HIV-1 replication.
- FEN1 activity facilitates the completion of double-stranded DNA synthesis for integrated provirus.
- FEN1 emerges as a potential therapeutic target for novel anti-HIV interventions.