Binding of human immunodeficiency virus type 1 (HIV-1) RNA to recombinant HIV-1 gag polyprotein - PubMed (original) (raw)

Binding of human immunodeficiency virus type 1 (HIV-1) RNA to recombinant HIV-1 gag polyprotein

J Luban et al. J Virol. 1991 Jun.

Abstract

We have expressed the human immunodeficiency virus type 1 (HIV-1) gag polyprotein (Pr55gag) in bacteria under the control of the T7 phage gene 10 promoter. When the gene encoding the viral protease is included in cis, in the -1 reading frame, the expected proteolytic cleavage products MA and CA are produced. Disruption of the protease-coding sequence prevents proteolytic processing, and full-length polyprotein is produced. Pr55gag, separated from bacterial proteins by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and immobilized on nitrocellulose membranes, binds RNA containing sequences from the 5' end of the HIV-1 genome. This binding is tolerant of a wide range of pH and temperature but has distinct salt preferences. Conditions were identified which prevented nonspecific binding of RNA to bacterial proteins but still allowed binding to Pr55gag. Under these conditions, irrelevant RNA probes lacking HIV-1 sequences bound Pr55gag less efficiently. Quantitation of binding to Pr55gag by HIV-1 RNA probes with deletions mutations demonstrated that there are two regions lying within the HIV-1 gag gene which independently promote binding of RNA to Pr55gag.

PubMed Disclaimer

Cited by

Basic residues in human immunodeficiency virus type 1 nucleocapsid promote virion assembly via interaction with RNA.
Cimarelli A, Sandin S, Höglund S, Luban J. Cimarelli A, et al. J Virol. 2000 Apr;74(7):3046-57. doi: 10.1128/jvi.74.7.3046-3057.2000. J Virol. 2000. PMID: 10708419 Free PMC article.
Sequence-specific interaction between HIV-1 matrix protein and viral genomic RNA revealed by in vitro genetic selection.
Purohit P, Dupont S, Stevenson M, Green MR. Purohit P, et al. RNA. 2001 Apr;7(4):576-84. doi: 10.1017/s1355838201002023. RNA. 2001. PMID: 11345436 Free PMC article.
In vitro selection of RNAs that bind to the human immunodeficiency virus type-1 gag polyprotein.
Lochrie MA, Waugh S, Pratt DG Jr, Clever J, Parslow TG, Polisky B. Lochrie MA, et al. Nucleic Acids Res. 1997 Jul 15;25(14):2902-10. doi: 10.1093/nar/25.14.2902. Nucleic Acids Res. 1997. PMID: 9207041 Free PMC article.
Determinants of the human immunodeficiency virus type 1 p15NC-RNA interaction that affect enhanced cleavage by the viral protease.
Sheng N, Pettit SC, Tritch RJ, Ozturk DH, Rayner MM, Swanstrom R, Erickson-Viitanen S. Sheng N, et al. J Virol. 1997 Aug;71(8):5723-32. doi: 10.1128/JVI.71.8.5723-5732.1997. J Virol. 1997. PMID: 9223458 Free PMC article.
Evidence for interstrand quadruplex formation in the dimerization of human immunodeficiency virus 1 genomic RNA.
Sundquist WI, Heaphy S. Sundquist WI, et al. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3393-7. doi: 10.1073/pnas.90.8.3393. Proc Natl Acad Sci U S A. 1993. PMID: 8475087 Free PMC article.

References

1. Nucleic Acids Res. 1990 Apr 11;18(7):1725-9 - PubMed
1. Genes Dev. 1990 Feb;4(2):277-86 - PubMed
1. J Virol. 1990 Jul;64(7):3207-11 - PubMed
1. J Virol. 1990 Oct;64(10):4978-87 - PubMed
1. J Virol. 1990 Oct;64(10):5230-4 - PubMed

Binding of human immunodeficiency virus type 1 (HIV-1) RNA to recombinant HIV-1 gag polyprotein - PubMed (original) (raw)