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. 1997 Aug;71(8):5723–5732. doi: 10.1128/jvi.71.8.5723-5732.1997

Determinants of the human immunodeficiency virus type 1 p15NC-RNA interaction that affect enhanced cleavage by the viral protease.

N Sheng 1, S C Pettit 1, R J Tritch 1, D H Ozturk 1, M M Rayner 1, R Swanstrom 1, S Erickson-Viitanen 1
PMCID: PMC191824  PMID: 9223458

Abstract

During human immunodeficiency virus type 1 (HIV-1) virion assembly, cleavage of the Gag precursor by the viral protease results in the transient appearance of a nucleocapsid-p1-p6 intermediate product designated p15NC. Utilizing the p15NC precursor protein produced with an in vitro transcription-translation system or purified after expression in Escherichia coli, we have demonstrated that RNA is required for efficient cleavage of HIV p15NC. Gel mobility shift and nitrocellulose filter binding experiments indicate that purified p15NC protein specifically binds its corresponding mRNA with an estimated Kd of 1.5 nM. Binding was not affected by the presence or absence of zinc or EDTA. Moreover, mutagenesis of the cysteine residues within either of the two Cys-His arrays had no effect on RNA binding or on RNA-dependent cleavage by the viral protease. In contrast, decreased binding of RNA and diminished susceptibility to cleavage in vitro were observed with p15NC-containing mutations in one or more residues within the triplet of basic amino acids present in the region between the two zinc fingers. In addition, we found that 21- to 24-base DNA and RNA oligonucleotides of a particular sequence and secondary structure could substitute for p15 RNA in the enhancement of p15NC cleavage. Virus particles carrying a mutation in the triplet of NC basic residues (P3BE) show delayed cleavage of p15NC and a defect in core formation despite the eventual appearance of fully processed virion protein. These results define determinants of the p15NC-RNA interaction that lead to enhanced protease-mediated cleavage and demonstrate the importance of the triplet of basic residues in formation of the virus core.

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Selected References

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  1. Aldovini A., Young R. A. Mutations of RNA and protein sequences involved in human immunodeficiency virus type 1 packaging result in production of noninfectious virus. J Virol. 1990 May;64(5):1920–1926. doi: 10.1128/jvi.64.5.1920-1926.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ashorn P., McQuade T. J., Thaisrivongs S., Tomasselli A. G., Tarpley W. G., Moss B. An inhibitor of the protease blocks maturation of human and simian immunodeficiency viruses and spread of infection. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7472–7476. doi: 10.1073/pnas.87.19.7472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barat C., Schatz O., Le Grice S., Darlix J. L. Analysis of the interactions of HIV1 replication primer tRNA(Lys,3) with nucleocapsid protein and reverse transcriptase. J Mol Biol. 1993 May 20;231(2):185–190. doi: 10.1006/jmbi.1993.1273. [DOI] [PubMed] [Google Scholar]
  4. Berkowitz R. D., Luban J., Goff S. P. Specific binding of human immunodeficiency virus type 1 gag polyprotein and nucleocapsid protein to viral RNAs detected by RNA mobility shift assays. J Virol. 1993 Dec;67(12):7190–7200. doi: 10.1128/jvi.67.12.7190-7200.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berkowitz R. D., Ohagen A., Höglund S., Goff S. P. Retroviral nucleocapsid domains mediate the specific recognition of genomic viral RNAs by chimeric Gag polyproteins during RNA packaging in vivo. J Virol. 1995 Oct;69(10):6445–6456. doi: 10.1128/jvi.69.10.6445-6456.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bowles N. E., Damay P., Spahr P. F. Effect of rearrangements and duplications of the Cys-His motifs of Rous sarcoma virus nucleocapsid protein. J Virol. 1993 Feb;67(2):623–631. doi: 10.1128/jvi.67.2.623-631.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cheng Y. S., McGowan M. H., Kettner C. A., Schloss J. V., Erickson-Viitanen S., Yin F. H. High-level synthesis of recombinant HIV-1 protease and the recovery of active enzyme from inclusion bodies. Gene. 1990 Mar 15;87(2):243–248. doi: 10.1016/0378-1119(90)90308-e. [DOI] [PubMed] [Google Scholar]
  8. Clever J., Sassetti C., Parslow T. G. RNA secondary structure and binding sites for gag gene products in the 5' packaging signal of human immunodeficiency virus type 1. J Virol. 1995 Apr;69(4):2101–2109. doi: 10.1128/jvi.69.4.2101-2109.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dannull J., Surovoy A., Jung G., Moelling K. Specific binding of HIV-1 nucleocapsid protein to PSI RNA in vitro requires N-terminal zinc finger and flanking basic amino acid residues. EMBO J. 1994 Apr 1;13(7):1525–1533. doi: 10.1002/j.1460-2075.1994.tb06414.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Darlix J. L., Lapadat-Tapolsky M., de Rocquigny H., Roques B. P. First glimpses at structure-function relationships of the nucleocapsid protein of retroviruses. J Mol Biol. 1995 Dec 8;254(4):523–537. doi: 10.1006/jmbi.1995.0635. [DOI] [PubMed] [Google Scholar]
  11. De Rocquigny H., Gabus C., Vincent A., Fournié-Zaluski M. C., Roques B., Darlix J. L. Viral RNA annealing activities of human immunodeficiency virus type 1 nucleocapsid protein require only peptide domains outside the zinc fingers. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6472–6476. doi: 10.1073/pnas.89.14.6472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dib-Hajj F., Khan R., Giedroc D. P. Retroviral nucleocapsid proteins possess potent nucleic acid strand renaturation activity. Protein Sci. 1993 Feb;2(2):231–243. doi: 10.1002/pro.5560020212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dorfman T., Luban J., Goff S. P., Haseltine W. A., Göttlinger H. G. Mapping of functionally important residues of a cysteine-histidine box in the human immunodeficiency virus type 1 nucleocapsid protein. J Virol. 1993 Oct;67(10):6159–6169. doi: 10.1128/jvi.67.10.6159-6169.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Draper D. E., Deckman I. C., Vartikar J. V. Physical studies of ribosomal protein-RNA interactions. Methods Enzymol. 1988;164:203–220. doi: 10.1016/s0076-6879(88)64044-4. [DOI] [PubMed] [Google Scholar]
  15. Erickson-Viitanen S., Manfredi J., Viitanen P., Tribe D. E., Tritch R., Hutchison C. A., 3rd, Loeb D. D., Swanstrom R. Cleavage of HIV-1 gag polyprotein synthesized in vitro: sequential cleavage by the viral protease. AIDS Res Hum Retroviruses. 1989 Dec;5(6):577–591. doi: 10.1089/aid.1989.5.577. [DOI] [PubMed] [Google Scholar]
  16. Feng Y. X., Copeland T. D., Henderson L. E., Gorelick R. J., Bosche W. J., Levin J. G., Rein A. HIV-1 nucleocapsid protein induces "maturation" of dimeric retroviral RNA in vitro. Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7577–7581. doi: 10.1073/pnas.93.15.7577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gelderblom H. R., Ozel M., Pauli G. Morphogenesis and morphology of HIV. Structure-function relations. Arch Virol. 1989;106(1-2):1–13. doi: 10.1007/BF01311033. [DOI] [PubMed] [Google Scholar]
  18. Gorelick R. J., Chabot D. J., Rein A., Henderson L. E., Arthur L. O. The two zinc fingers in the human immunodeficiency virus type 1 nucleocapsid protein are not functionally equivalent. J Virol. 1993 Jul;67(7):4027–4036. doi: 10.1128/jvi.67.7.4027-4036.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gorelick R. J., Henderson L. E., Hanser J. P., Rein A. Point mutants of Moloney murine leukemia virus that fail to package viral RNA: evidence for specific RNA recognition by a "zinc finger-like" protein sequence. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8420–8424. doi: 10.1073/pnas.85.22.8420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gorelick R. J., Nigida S. M., Jr, Bess J. W., Jr, Arthur L. O., Henderson L. E., Rein A. Noninfectious human immunodeficiency virus type 1 mutants deficient in genomic RNA. J Virol. 1990 Jul;64(7):3207–3211. doi: 10.1128/jvi.64.7.3207-3211.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gowda S. D., Stein B. S., Engleman E. G. Identification of protein intermediates in the processing of the p55 HIV-1 gag precursor in cells infected with recombinant vaccinia virus. J Biol Chem. 1989 May 25;264(15):8459–8462. [PubMed] [Google Scholar]
  22. Henderson L. E., Sowder R. C., Copeland T. D., Oroszlan S., Benveniste R. E. Gag precursors of HIV and SIV are cleaved into six proteins found in the mature virions. J Med Primatol. 1990;19(3-4):411–419. [PubMed] [Google Scholar]
  23. Housset V., De Rocquigny H., Roques B. P., Darlix J. L. Basic amino acids flanking the zinc finger of Moloney murine leukemia virus nucleocapsid protein NCp10 are critical for virus infectivity. J Virol. 1993 May;67(5):2537–2545. doi: 10.1128/jvi.67.5.2537-2545.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jamjoom G. A., Naso R. B., Arlinghaus R. B. Selective decrease in the rate of cleavage of an intracellular precursor to Rauscher leukemia virus p30 by treatment of infected cells with actinomycin D. J Virol. 1976 Sep;19(3):1054–1072. doi: 10.1128/jvi.19.3.1054-1072.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jowett J. B., Hockley D. J., Nermut M. V., Jones I. M. Distinct signals in human immunodeficiency virus type 1 Pr55 necessary for RNA binding and particle formation. J Gen Virol. 1992 Dec;73(Pt 12):3079–3086. doi: 10.1099/0022-1317-73-12-3079. [DOI] [PubMed] [Google Scholar]
  26. Kageyama S., Hoekzema D. T., Murakawa Y., Kojima E., Shirasaka T., Kempf D. J., Norbeck D. W., Erickson J., Mitsuya H. A C2 symmetry-based HIV protease inhibitor, A77003, irreversibly inhibits infectivity of HIV-1 in vitro. AIDS Res Hum Retroviruses. 1994 Jun;10(6):735–743. doi: 10.1089/aid.1994.10.735. [DOI] [PubMed] [Google Scholar]
  27. Kaplan A. H., Manchester M., Everitt L., Swanstrom R. Analysis of human immunodeficiency virus type 1 protease activity in eukaryotic and bacterial cells. Methods Enzymol. 1994;241:58–69. doi: 10.1016/0076-6879(94)41059-3. [DOI] [PubMed] [Google Scholar]
  28. Kaplan A. H., Zack J. A., Knigge M., Paul D. A., Kempf D. J., Norbeck D. W., Swanstrom R. Partial inhibition of the human immunodeficiency virus type 1 protease results in aberrant virus assembly and the formation of noninfectious particles. J Virol. 1993 Jul;67(7):4050–4055. doi: 10.1128/jvi.67.7.4050-4055.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Khan R., Giedroc D. P. Nucleic acid binding properties of recombinant Zn2 HIV-1 nucleocapsid protein are modulated by COOH-terminal processing. J Biol Chem. 1994 Sep 9;269(36):22538–22546. [PubMed] [Google Scholar]
  30. Kohl N. E., Emini E. A., Schleif W. A., Davis L. J., Heimbach J. C., Dixon R. A., Scolnick E. M., Sigal I. S. Active human immunodeficiency virus protease is required for viral infectivity. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4686–4690. doi: 10.1073/pnas.85.13.4686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kräusslich H. G., Fäcke M., Heuser A. M., Konvalinka J., Zentgraf H. The spacer peptide between human immunodeficiency virus capsid and nucleocapsid proteins is essential for ordered assembly and viral infectivity. J Virol. 1995 Jun;69(6):3407–3419. doi: 10.1128/jvi.69.6.3407-3419.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  33. Lam W. C., Maki A. H., Casas-Finet J. R., Erickson J. W., Kane B. P., Sowder R. C., 2nd, Henderson L. E. Phosphorescence and optically detected magnetic resonance investigation of the binding of the nucleocapsid protein of the human immunodeficiency virus type 1 and related peptides to RNA. Biochemistry. 1994 Sep 6;33(35):10693–10700. doi: 10.1021/bi00201a017. [DOI] [PubMed] [Google Scholar]
  34. Lambert D. M., Petteway S. R., Jr, McDanal C. E., Hart T. K., Leary J. J., Dreyer G. B., Meek T. D., Bugelski P. J., Bolognesi D. P., Metcalf B. W. Human immunodeficiency virus type 1 protease inhibitors irreversibly block infectivity of purified virions from chronically infected cells. Antimicrob Agents Chemother. 1992 May;36(5):982–988. doi: 10.1128/aac.36.5.982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Li M. S., Garcia-Asua G., Bhattacharyya U., Mascagni P., Austen B. M., Roberts M. M. The Vpr protein of human immunodeficiency virus type 1 binds to nucleocapsid protein p7 in vitro. Biochem Biophys Res Commun. 1996 Jan 5;218(1):352–355. doi: 10.1006/bbrc.1996.0061. [DOI] [PubMed] [Google Scholar]
  36. Li X., Gu Z., Geleziunas R., Kleiman L., Wainberg M. A., Parniak M. A. Expression, purification, and RNA-binding properties of HIV-1 p15gag nucleocapsid protein. Protein Expr Purif. 1993 Aug;4(4):304–311. doi: 10.1006/prep.1993.1039. [DOI] [PubMed] [Google Scholar]
  37. Luban J., Goff S. P. Binding of human immunodeficiency virus type 1 (HIV-1) RNA to recombinant HIV-1 gag polyprotein. J Virol. 1991 Jun;65(6):3203–3212. doi: 10.1128/jvi.65.6.3203-3212.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. McBride M. S., Panganiban A. T. The human immunodeficiency virus type 1 encapsidation site is a multipartite RNA element composed of functional hairpin structures. J Virol. 1996 May;70(5):2963–2973. doi: 10.1128/jvi.70.5.2963-2973.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Mervis R. J., Ahmad N., Lillehoj E. P., Raum M. G., Salazar F. H., Chan H. W., Venkatesan S. The gag gene products of human immunodeficiency virus type 1: alignment within the gag open reading frame, identification of posttranslational modifications, and evidence for alternative gag precursors. J Virol. 1988 Nov;62(11):3993–4002. doi: 10.1128/jvi.62.11.3993-4002.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Morellet N., Jullian N., De Rocquigny H., Maigret B., Darlix J. L., Roques B. P. Determination of the structure of the nucleocapsid protein NCp7 from the human immunodeficiency virus type 1 by 1H NMR. EMBO J. 1992 Aug;11(8):3059–3065. doi: 10.1002/j.1460-2075.1992.tb05377.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Morellet N., de Rocquigny H., Mély Y., Jullian N., Déméné H., Ottmann M., Gérard D., Darlix J. L., Fournie-Zaluski M. C., Roques B. P. Conformational behaviour of the active and inactive forms of the nucleocapsid NCp7 of HIV-1 studied by 1H NMR. J Mol Biol. 1994 Jan 7;235(1):287–301. doi: 10.1016/s0022-2836(05)80033-6. [DOI] [PubMed] [Google Scholar]
  42. Mély Y., de Rocquigny H., Sorinas-Jimeno M., Keith G., Roques B. P., Marquet R., Gérard D. Binding of the HIV-1 nucleocapsid protein to the primer tRNA(3Lys), in vitro, is essentially not specific. J Biol Chem. 1995 Jan 27;270(4):1650–1656. doi: 10.1074/jbc.270.4.1650. [DOI] [PubMed] [Google Scholar]
  43. Oroszlan S., Luftig R. B. Retroviral proteinases. Curr Top Microbiol Immunol. 1990;157:153–185. doi: 10.1007/978-3-642-75218-6_6. [DOI] [PubMed] [Google Scholar]
  44. Ottmann M., Gabus C., Darlix J. L. The central globular domain of the nucleocapsid protein of human immunodeficiency virus type 1 is critical for virion structure and infectivity. J Virol. 1995 Mar;69(3):1778–1784. doi: 10.1128/jvi.69.3.1778-1784.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Pettit S. C., Moody M. D., Wehbie R. S., Kaplan A. H., Nantermet P. V., Klein C. A., Swanstrom R. The p2 domain of human immunodeficiency virus type 1 Gag regulates sequential proteolytic processing and is required to produce fully infectious virions. J Virol. 1994 Dec;68(12):8017–8027. doi: 10.1128/jvi.68.12.8017-8027.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Poon D. T., Wu J., Aldovini A. Charged amino acid residues of human immunodeficiency virus type 1 nucleocapsid p7 protein involved in RNA packaging and infectivity. J Virol. 1996 Oct;70(10):6607–6616. doi: 10.1128/jvi.70.10.6607-6616.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Ratner L., Fisher A., Jagodzinski L. L., Mitsuya H., Liou R. S., Gallo R. C., Wong-Staal F. Complete nucleotide sequences of functional clones of the AIDS virus. AIDS Res Hum Retroviruses. 1987 Spring;3(1):57–69. doi: 10.1089/aid.1987.3.57. [DOI] [PubMed] [Google Scholar]
  48. Rayner M. M., Cordova B. C., Meade R. P., Aldrich P. E., Jadhav P. K., Ru Y., Lam P. Y. DMP 323, a nonpeptide cyclic urea inhibitor of human immunodeficiency virus (HIV) protease, specifically and persistently blocks intracellular processing of HIV gag polyprotein. Antimicrob Agents Chemother. 1994 Jul;38(7):1635–1640. doi: 10.1128/aac.38.7.1635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Reicin A. S., Paik S., Berkowitz R. D., Luban J., Lowy I., Goff S. P. Linker insertion mutations in the human immunodeficiency virus type 1 gag gene: effects on virion particle assembly, release, and infectivity. J Virol. 1995 Feb;69(2):642–650. doi: 10.1128/jvi.69.2.642-650.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Rice W. G., Turpin J. A., Schaeffer C. A., Graham L., Clanton D., Buckheit R. W., Jr, Zaharevitz D., Summers M. F., Wallqvist A., Covell D. G. Evaluation of selected chemotypes in coupled cellular and molecular target-based screens identifies novel HIV-1 zinc finger inhibitors. J Med Chem. 1996 Sep 13;39(19):3606–3616. doi: 10.1021/jm960375o. [DOI] [PubMed] [Google Scholar]
  51. Rosenberg A. H., Lade B. N., Chui D. S., Lin S. W., Dunn J. J., Studier F. W. Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene. 1987;56(1):125–135. doi: 10.1016/0378-1119(87)90165-x. [DOI] [PubMed] [Google Scholar]
  52. Ross E. K., Fuerst T. R., Orenstein J. M., O'Neill T., Martin M. A., Venkatesan S. Maturation of human immunodeficiency virus particles assembled from the gag precursor protein requires in situ processing by gag-pol protease. AIDS Res Hum Retroviruses. 1991 May;7(5):475–483. doi: 10.1089/aid.1991.7.475. [DOI] [PubMed] [Google Scholar]
  53. Sakaguchi K., Zambrano N., Baldwin E. T., Shapiro B. A., Erickson J. W., Omichinski J. G., Clore G. M., Gronenborn A. M., Appella E. Identification of a binding site for the human immunodeficiency virus type 1 nucleocapsid protein. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5219–5223. doi: 10.1073/pnas.90.11.5219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Schmalzbauer E., Strack B., Dannull J., Guehmann S., Moelling K. Mutations of basic amino acids of NCp7 of human immunodeficiency virus type 1 affect RNA binding in vitro. J Virol. 1996 Feb;70(2):771–777. doi: 10.1128/jvi.70.2.771-777.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Schätzl H., Gelderblom H. R., Nitschko H., von der Helm K. Analysis of non-infectious HIV particles produced in presence of HIV proteinase inhibitor. Arch Virol. 1991;120(1-2):71–81. doi: 10.1007/BF01310950. [DOI] [PubMed] [Google Scholar]
  56. Sheng N., Erickson-Viitanen S. Cleavage of p15 protein in vitro by human immunodeficiency virus type 1 protease is RNA dependent. J Virol. 1994 Oct;68(10):6207–6214. doi: 10.1128/jvi.68.10.6207-6214.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. South T. L., Summers M. F. Zinc- and sequence-dependent binding to nucleic acids by the N-terminal zinc finger of the HIV-1 nucleocapsid protein: NMR structure of the complex with the Psi-site analog, dACGCC. Protein Sci. 1993 Jan;2(1):3–19. doi: 10.1002/pro.5560020102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Surovoy A., Dannull J., Moelling K., Jung G. Conformational and nucleic acid binding studies on the synthetic nucleocapsid protein of HIV-1. J Mol Biol. 1993 Jan 5;229(1):94–104. doi: 10.1006/jmbi.1993.1011. [DOI] [PubMed] [Google Scholar]
  59. Tanchou V., Gabus C., Rogemond V., Darlix J. L. Formation of stable and functional HIV-1 nucleoprotein complexes in vitro. J Mol Biol. 1995 Oct 6;252(5):563–571. doi: 10.1006/jmbi.1995.0520. [DOI] [PubMed] [Google Scholar]
  60. Tritch R. J., Cheng Y. E., Yin F. H., Erickson-Viitanen S. Mutagenesis of protease cleavage sites in the human immunodeficiency virus type 1 gag polyprotein. J Virol. 1991 Feb;65(2):922–930. doi: 10.1128/jvi.65.2.922-930.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Turner D. H., Sugimoto N., Freier S. M. RNA structure prediction. Annu Rev Biophys Biophys Chem. 1988;17:167–192. doi: 10.1146/annurev.bb.17.060188.001123. [DOI] [PubMed] [Google Scholar]
  62. Turpin J. A., Terpening S. J., Schaeffer C. A., Yu G., Glover C. J., Felsted R. L., Sausville E. A., Rice W. G. Inhibitors of human immunodeficiency virus type 1 zinc fingers prevent normal processing of gag precursors and result in the release of noninfectious virus particles. J Virol. 1996 Sep;70(9):6180–6189. doi: 10.1128/jvi.70.9.6180-6189.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Veronese F. D., Rahman R., Copeland T. D., Oroszlan S., Gallo R. C., Sarngadharan M. G. Immunological and chemical analysis of P6, the carboxyl-terminal fragment of HIV P15. AIDS Res Hum Retroviruses. 1987 Fall;3(3):253–264. doi: 10.1089/aid.1987.3.253. [DOI] [PubMed] [Google Scholar]
  64. Weiss S., König B., Morikawa Y., Jones I. Recombinant HIV-1 nucleocapsid protein p15 produced as a fusion protein with glutathione S-transferase in Escherichia coli mediates dimerization and enhances reverse transcription of retroviral RNA. Gene. 1992 Nov 16;121(2):203–212. doi: 10.1016/0378-1119(92)90123-7. [DOI] [PubMed] [Google Scholar]
  65. Wu W., Henderson L. E., Copeland T. D., Gorelick R. J., Bosche W. J., Rein A., Levin J. G. Human immunodeficiency virus type 1 nucleocapsid protein reduces reverse transcriptase pausing at a secondary structure near the murine leukemia virus polypurine tract. J Virol. 1996 Oct;70(10):7132–7142. doi: 10.1128/jvi.70.10.7132-7142.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. You J. C., McHenry C. S. HIV nucleocapsid protein. Expression in Escherichia coli, purification, and characterization. J Biol Chem. 1993 Aug 5;268(22):16519–16527. [PubMed] [Google Scholar]
  67. Zhang Y., Barklis E. Nucleocapsid protein effects on the specificity of retrovirus RNA encapsidation. J Virol. 1995 Sep;69(9):5716–5722. doi: 10.1128/jvi.69.9.5716-5722.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  68. Zuker M. On finding all suboptimal foldings of an RNA molecule. Science. 1989 Apr 7;244(4900):48–52. doi: 10.1126/science.2468181. [DOI] [PubMed] [Google Scholar]

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