Molecular characterization of proteolytic processing of the Gag proteins of human spumavirus.

Abstract:

:Spumaviruses, or foamy viruses, express Gag proteins that are incompletely processed by the viral protease in cell cultures. To delineate the proteolytic cleavage sites between potential Gag subdomains, recombinant human spumaretrovirus (HSRV) Gag proteins of different lengths were expressed, purified by affinity chromatography, and subjected to HSRV protease assays. HSRV-specific proteolytic cleavage products were isolated and characterized by Western blotting. Peptides spanning potential cleavage sites, as deduced from the sizes of the proteolytic cleavage products, were chemically synthesized and assayed with HSRV protease. The cleaved peptides were then subjected to mass spectrometry. In control experiments, HSRV protease-deficient mutant proteins were used to rule out unspecific processing by nonviral proteases. The cleavage site junctions identified and the calculated sizes of the cleavage products were in agreement with those of the authentic cleavage products of the HSRV Gag proteins detectable in viral proteins from purified HSRV particles and in virus-infected cells. The biological significance of the data was confirmed by mutational analysis of the cleavage sites in a recombinant Gag protein and in the context of the infectious HSRV DNA provirus.

journal_name

J Virol

journal_title

Journal of virology

authors

Pfrepper KI,Löchelt M,Rackwitz HR,Schnölzer M,Heid H,Flügel RM

doi

10.1128/JVI.73.9.7907-7911.1999

subject

Has Abstract

pub_date

1999-09-01 00:00:00

pages

7907-11

issue

9

eissn

0022-538X

issn

1098-5514

journal_volume

73

pub_type

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