DOI: 10.1073/pnas.0401312101. Reconstitution of retroviral fusion and uncoating in a cell-free system

Reconstitution of retroviral fusion and uncoating in a cell-free system

10.1073/pnas.0401312101

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The molecular events underlying the immediate steps of retroviral uncoating, occurring after membrane fusion and leading to the formation of an active reverse transcription complex, are not known. To better understand these processes, we have developed a cell-free system that recapitulates these early steps of retroviral replication by using avian sarcoma and leukosis virus as a model retrovirus. The substrates used in this system are viral particles that are trapped before completing membrane fusion. These virions are induced to fuse out of endosomes and the viral cores are released into solution where they are amenable to biochemical manipulation. This system revealed that membrane fusion is not sufficient to stimulate the formation of a reverse transcription complex. Instead, ATP hydrolysis and cellular factors >5 kDa in size are required. Furthermore, later steps of avian sarcoma and leukosis virus reverse transcription were stimulated by nuclear factors. The cell-free system should now allow for the definition of retroviral uncoating mechanisms and facilitate the identification and characterization of the cellular factors involved.

Bibliography

Narayan, S., & Young, J. A. T. (2004). Reconstitution of retroviral fusion and uncoating in a cell-free system. Proceedings of the National Academy of Sciences, 101(20), 7721â7726.

Authors 2

Shakti Narayan (first)
John A. T. Young (additional)

References 42 Referenced 22

10.1146/annurev.biochem.70.1.777
Barnard, R. J. & Young, J. A. T. (2003) Curr. Top. Microbiol. 281, 107-136. / Curr. Top. Microbiol. (2003)
Pierson, T. C. & Doms, R. W. (2003) Curr. Top. Microbiol. 281, 1-28. / Curr. Top. Microbiol. (2003)
Dvorin, J. D. & Malim, M. H. (2003) Curr. Top. Microbiol. 281, 179-208. / Curr. Top. Microbiol. (2003)
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Ausubel F. M. Brent R. Kingston R. E. Moore D. D. Seidman J. G. Smith J. A. & Struhl K. (2000) Current Protocols in Molecular Biology (Wiley New York) Vol. 1 . (10.1002/0471142727)
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10.1146/annurev.cellbio.16.1.627

Dates

Type	When
Created	21 years, 3 months ago (May 5, 2004, 8:13 p.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 9:46 a.m.)
Indexed	1 year, 3 months ago (May 17, 2024, 3:43 p.m.)
Issued	21 years, 3 months ago (May 5, 2004)
Published	21 years, 3 months ago (May 5, 2004)
Published Online	21 years, 3 months ago (May 5, 2004)
Published Print	21 years, 3 months ago (May 18, 2004)

Funders 0

None

BibTeX

@article{Narayan_2004, title={Reconstitution of retroviral fusion and uncoating in a cell-free system}, volume={101}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0401312101}, DOI={10.1073/pnas.0401312101}, number={20}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Narayan, Shakti and Young, John A. T.}, year={2004}, month=may, pages={7721–7726} }

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