Multiple barriers to recombination between divergent HIV-1 variants revealed by a dual-marker recombination assay
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Standard
Multiple barriers to recombination between divergent HIV-1 variants revealed by a dual-marker recombination assay. / Nikolaitchik, Olga A.; Galli, Andrea; Moore, Michael D.; Pathak, Vinay K.; Hu, Wei Shau.
I: Journal of Molecular Biology, Bind 407, Nr. 4, 08.04.2011, s. 521-531.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Multiple barriers to recombination between divergent HIV-1 variants revealed by a dual-marker recombination assay
AU - Nikolaitchik, Olga A.
AU - Galli, Andrea
AU - Moore, Michael D.
AU - Pathak, Vinay K.
AU - Hu, Wei Shau
N1 - Funding Information: We thank Hans-Georg Krausslich for group O molecular clone. This research was supported by the Intramural Research Program of the National Institutes of Health, National Cancer Institute, Center for Cancer Research.
PY - 2011/4/8
Y1 - 2011/4/8
N2 - Recombination is a major force for generating human immunodeficiency virus type 1 (HIV-1) diversity and produces numerous recombinants circulating in the human population. We previously established a cell-based system using green fluorescent protein gene (gfp) as a reporter to study the mechanisms of HIV-1 recombination. We now report an improved system capable of detecting recombination using authentic viral sequences. Frameshift mutations were introduced into the gag gene so that parental viruses do not express full-length Gag; however, recombination can generate a progeny virus that expresses a functional Gag. We demonstrate that this Gag reconstitution assay can be used to detect recombination between two group M HIV-1 variants of the same or of different subtypes. Using both gfp and gag assays, we found that, similar to group M viruses, group O viruses also recombine frequently. When recombination between a group M virus and a group O virus was examined, we found three distinct barriers for intergroup recombination. First, similar to recombination within group M viruses, intergroup recombination is affected by the identity of the dimerization initiation signal (DIS); variants with the same DIS recombined at a higher rate than those with different DIS. Second, using the gfp recombination assay, we showed that intergroup recombination occurs much less frequently than intragroup recombination, even though the gfp target sequence is identical in all viruses. Finally, Gag reconstitution between variants from different groups is further reduced compared with green fluorescent protein, indicating that sequence divergence interferes with recombination efficiency in the gag gene. Compared with identical sequences, we estimate that recombination rates are reduced by 3-fold and by 10- to 13-fold when the target regions in gag contain 91% and 72-73% sequence identities, respectively. These results show that there are at least three distinct mechanisms preventing exchange of genetic information between divergent HIV-1 variants from different groups.
AB - Recombination is a major force for generating human immunodeficiency virus type 1 (HIV-1) diversity and produces numerous recombinants circulating in the human population. We previously established a cell-based system using green fluorescent protein gene (gfp) as a reporter to study the mechanisms of HIV-1 recombination. We now report an improved system capable of detecting recombination using authentic viral sequences. Frameshift mutations were introduced into the gag gene so that parental viruses do not express full-length Gag; however, recombination can generate a progeny virus that expresses a functional Gag. We demonstrate that this Gag reconstitution assay can be used to detect recombination between two group M HIV-1 variants of the same or of different subtypes. Using both gfp and gag assays, we found that, similar to group M viruses, group O viruses also recombine frequently. When recombination between a group M virus and a group O virus was examined, we found three distinct barriers for intergroup recombination. First, similar to recombination within group M viruses, intergroup recombination is affected by the identity of the dimerization initiation signal (DIS); variants with the same DIS recombined at a higher rate than those with different DIS. Second, using the gfp recombination assay, we showed that intergroup recombination occurs much less frequently than intragroup recombination, even though the gfp target sequence is identical in all viruses. Finally, Gag reconstitution between variants from different groups is further reduced compared with green fluorescent protein, indicating that sequence divergence interferes with recombination efficiency in the gag gene. Compared with identical sequences, we estimate that recombination rates are reduced by 3-fold and by 10- to 13-fold when the target regions in gag contain 91% and 72-73% sequence identities, respectively. These results show that there are at least three distinct mechanisms preventing exchange of genetic information between divergent HIV-1 variants from different groups.
KW - CA
KW - capsid
KW - dimerization initiation signal
KW - DIS
KW - GFP
KW - green fluorescent protein
KW - heat-stable antigen
KW - HIV-1
KW - HSA
KW - human immunodeficiency virus type 1
KW - internal ribosome entry site
KW - IRES
KW - long terminal repeat
KW - LTR
KW - MOI
KW - multiplicity of infection
KW - PE
KW - phycoerythrin
KW - reverse transcriptase
KW - RT
KW - simian immunodeficiency virus
KW - SIV
UR - http://www.scopus.com/inward/record.url?scp=79952438912&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2011.01.052
DO - 10.1016/j.jmb.2011.01.052
M3 - Journal article
C2 - 21295586
AN - SCOPUS:79952438912
VL - 407
SP - 521
EP - 531
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 4
ER -
ID: 379291849