Retrovirus-mediated insertion of expressed and non-expressed genes at identical chromosomal locations

Brent Berwin, Eric Barklis

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

During retrovirus replication, a cellularly derived tRNA is annealed to the viral RNA at the primer binding site (PBS) to prime reverse transcription, and both the tRNA and the PBS become copied and matched together on complementary proviral DNA strands prior to integration. Using a viral PBS single base pair mutant which affects provirus expression in undifferentiated cells, we show that reversion to wild type (wt) occurs at a frequency of approximately 50%. Daughter cell lines containing wt or mutant proviruses at identical chromosomal sites have been isolated, supporting a model where an integrated PBS-mismatched provirus was copied before mismatch correction could occur. Virus expression in daughter cells containing the mutant provirus was 100-fold higher than in cells bearing the wt counterpart. Additionally, proviral 5′ DNA and cellular 5′ flanking DNA became methylated in daughter cells containing wt but not mutant integrants. These results strongly support the current model of retrovirus reverse transcription, and indicate that the wt PBS region contains an element which suppresses virus expression and directs the methylation of viral and neighboring cellular DNA.

Original languageEnglish (US)
Pages (from-to)2399-2407
Number of pages9
JournalNucleic Acids Research
Volume21
Issue number10
StatePublished - May 25 1993

Fingerprint

Binding sites
Retroviridae
Proviruses
Insertion
Mutant
Genes
Binding Sites
Gene
DNA
Cell
Transcription
Transfer RNA
Viruses
Reverse Transcription
Virus
Reverse
Bearings (structural)
Methylation
Viral RNA
RNA

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Retrovirus-mediated insertion of expressed and non-expressed genes at identical chromosomal locations. / Berwin, Brent; Barklis, Eric.

In: Nucleic Acids Research, Vol. 21, No. 10, 25.05.1993, p. 2399-2407.

Research output: Contribution to journalArticle

@article{4c29b27467cb42f8aac284704b7d9b9c,
title = "Retrovirus-mediated insertion of expressed and non-expressed genes at identical chromosomal locations",
abstract = "During retrovirus replication, a cellularly derived tRNA is annealed to the viral RNA at the primer binding site (PBS) to prime reverse transcription, and both the tRNA and the PBS become copied and matched together on complementary proviral DNA strands prior to integration. Using a viral PBS single base pair mutant which affects provirus expression in undifferentiated cells, we show that reversion to wild type (wt) occurs at a frequency of approximately 50{\%}. Daughter cell lines containing wt or mutant proviruses at identical chromosomal sites have been isolated, supporting a model where an integrated PBS-mismatched provirus was copied before mismatch correction could occur. Virus expression in daughter cells containing the mutant provirus was 100-fold higher than in cells bearing the wt counterpart. Additionally, proviral 5′ DNA and cellular 5′ flanking DNA became methylated in daughter cells containing wt but not mutant integrants. These results strongly support the current model of retrovirus reverse transcription, and indicate that the wt PBS region contains an element which suppresses virus expression and directs the methylation of viral and neighboring cellular DNA.",
author = "Brent Berwin and Eric Barklis",
year = "1993",
month = "5",
day = "25",
language = "English (US)",
volume = "21",
pages = "2399--2407",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "10",

}

TY - JOUR

T1 - Retrovirus-mediated insertion of expressed and non-expressed genes at identical chromosomal locations

AU - Berwin, Brent

AU - Barklis, Eric

PY - 1993/5/25

Y1 - 1993/5/25

N2 - During retrovirus replication, a cellularly derived tRNA is annealed to the viral RNA at the primer binding site (PBS) to prime reverse transcription, and both the tRNA and the PBS become copied and matched together on complementary proviral DNA strands prior to integration. Using a viral PBS single base pair mutant which affects provirus expression in undifferentiated cells, we show that reversion to wild type (wt) occurs at a frequency of approximately 50%. Daughter cell lines containing wt or mutant proviruses at identical chromosomal sites have been isolated, supporting a model where an integrated PBS-mismatched provirus was copied before mismatch correction could occur. Virus expression in daughter cells containing the mutant provirus was 100-fold higher than in cells bearing the wt counterpart. Additionally, proviral 5′ DNA and cellular 5′ flanking DNA became methylated in daughter cells containing wt but not mutant integrants. These results strongly support the current model of retrovirus reverse transcription, and indicate that the wt PBS region contains an element which suppresses virus expression and directs the methylation of viral and neighboring cellular DNA.

AB - During retrovirus replication, a cellularly derived tRNA is annealed to the viral RNA at the primer binding site (PBS) to prime reverse transcription, and both the tRNA and the PBS become copied and matched together on complementary proviral DNA strands prior to integration. Using a viral PBS single base pair mutant which affects provirus expression in undifferentiated cells, we show that reversion to wild type (wt) occurs at a frequency of approximately 50%. Daughter cell lines containing wt or mutant proviruses at identical chromosomal sites have been isolated, supporting a model where an integrated PBS-mismatched provirus was copied before mismatch correction could occur. Virus expression in daughter cells containing the mutant provirus was 100-fold higher than in cells bearing the wt counterpart. Additionally, proviral 5′ DNA and cellular 5′ flanking DNA became methylated in daughter cells containing wt but not mutant integrants. These results strongly support the current model of retrovirus reverse transcription, and indicate that the wt PBS region contains an element which suppresses virus expression and directs the methylation of viral and neighboring cellular DNA.

UR - http://www.scopus.com/inward/record.url?scp=0027287820&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027287820&partnerID=8YFLogxK

M3 - Article

C2 - 8506135

AN - SCOPUS:0027287820

VL - 21

SP - 2399

EP - 2407

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 10

ER -