Ty1 proteolytic cleavage sites are required for transposition

All sites are not created equal

G. V. Merkulov, Jr Lawler J.F., Y. Eby, J. D. Boeke

Research output: Contribution to journalArticle

Abstract

The retroviral protease is a key enzyme in a viral multienzyme complex that initiates an ordered sequence of events leading to virus assembly and propagation. Viral peptides are initially synthesized as polyprotein precursors; these precursors undergo a number of proteolytic cleavages executed by the protease in a specific and presumably ordered manner. To determine the role of individual protease cleavage sites in Ty1, a retrotransposon from Saccharomyces cerevisiae, the cleavage sites were systematically mutagenized. Altering the cleavage sites of the yeast Ty1 retrotransposon produces mutants with distinct retrotransposition phenotypes. Blocking the Gag/PR site also blocks cleavage at the other two cleavage sites, PR/IN and IN/RT. In contrast, mutational block of the PR/IN or IN/RT sites does not prevent cleavage at the other two sites. Retrotransposons with mutations in each of these sites have transposition defects. Mutations in the PR/IN and IN/RT sites, but not in the Gag/PR site, can be complemented in trans by endogenous Ty1 copies. Hence, the digestion of the Gag/PR site and release of the protease N terminus is a prerequisite for processing at the remaining sites; cleavage of PR/IN is not required for the cleavage of IN/RT, and vice versa. Of the three cleavage sites in the Gag-Pol precursor, the Gag/PR site is processed first. Thus, Ty1 Gag-Pol processing proceeds by an ordered pathway.

Original languageEnglish (US)
Pages (from-to)638-644
Number of pages7
JournalJournal of Virology
Volume75
Issue number2
DOIs
StatePublished - 2001

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transposition (genetics)
Retroelements
retrotransposons
proteinases
Peptide Hydrolases
multienzyme complexes
Multienzyme Complexes
mutation
Virus Assembly
Polyproteins
Mutation
Saccharomyces cerevisiae
Digestion
Yeasts
digestion
peptides
yeasts
Phenotype
phenotype
mutants

ASJC Scopus subject areas

  • Immunology

Cite this

Ty1 proteolytic cleavage sites are required for transposition : All sites are not created equal. / Merkulov, G. V.; Lawler J.F., Jr; Eby, Y.; Boeke, J. D.

In: Journal of Virology, Vol. 75, No. 2, 2001, p. 638-644.

Research output: Contribution to journalArticle

Merkulov, G. V. ; Lawler J.F., Jr ; Eby, Y. ; Boeke, J. D. / Ty1 proteolytic cleavage sites are required for transposition : All sites are not created equal. In: Journal of Virology. 2001 ; Vol. 75, No. 2. pp. 638-644.
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