HIV-1 nucleocapsid protein zinc finger structures induce tRNALys,3 structural changes but are not critical for primer/template annealing11Edited by M. F. Summers

Hargittai, Michele R.S.; Mangla, Anil T.; Gorelick, Robert J.; Musier‐Forsyth, Karin

doi:10.1006/jmbi.2001.5021

Cited by 84 publications

(109 citation statements)

References 73 publications

(104 reference statements)

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“…It is possible that there is only one activity that stimulates hybridization and then unwinds secondary structure through strand invasion (71). However, others have clearly shown that NC destabilizes secondary structure in tRNAs and other nucleic acids in the absence of a complementary strand (37,72). This provides clear evidence for unwinding activity, whereas the enhanced hybridization in the absence of structure shown here supports a separate annealing activity.…”

Section: Mutant Nc Proteins Display Differing Effects On Annealing Dmentioning

confidence: 52%

Differing Roles of the N- and C-terminal Zinc Fingers in Human Immunodeficiency Virus Nucleocapsid Protein-enhanced Nucleic Acid Annealing

Heath

Derebail

Gorelick

et al. 2003

Journal of Biological Chemistry

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The replication process of human immunodeficiency virus requires a number of nucleic acid annealing steps facilitated by the hybridization and helix-destabilizing activities of human immunodeficiency virus nucleocapsid (NC) protein. NC contains two CCHC zinc finger motifs numbered 1 and 2 from the N terminus. The amino acids surrounding the CCHC residues differ between the two zinc fingers. Assays were preformed to investigate the activities of the fingers by determining the effect of mutant and wild-type proteins on annealing of 42-nucleotide RNA and DNA complements. The mutants 1.1 NC and 2.2 NC had duplications of the N-and C-terminal zinc fingers in positions 1 and 2. The mutant 2.1 NC had the native zinc fingers with their positions switched. Annealing assays were completed with unstructured and highly structured oligonucleotide complements. 2.2 NC had a near wild-type level of annealing of unstructured nucleic acids, whereas it was completely unable to stimulate annealing of highly structured nucleic acids. In contrast, 1.1 NC was able to stimulate annealing of both unstructured and structured substrates, but to a lesser degree than the wild-type protein. Results suggest that finger 1 has a greater role in unfolding of strong secondary structures, whereas finger 2 serves an accessory role that leads to a further increase in the rate of annealing.

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Section: Mutant Nc Proteins Display Differing Effects On Annealing Dmentioning

confidence: 52%

Differing Roles of the N- and C-terminal Zinc Fingers in Human Immunodeficiency Virus Nucleocapsid Protein-enhanced Nucleic Acid Annealing

Heath

Derebail

Gorelick

et al. 2003

Journal of Biological Chemistry

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“…In HIV-1, mature nucleocapsid protein NCp7 facilitates the in vitro annealing of tRNA Lys3 to in vitro transcribed genomic RNA sequence (43), probably by unwinding the secondary structure of stem/loops in the PBS area of the genomic RNA (31). It has been demonstrated by several groups that nucleocapsid protein does not unwind the secondary structure of tRNA in vitro, and the protein only has very subtle tertiary structural and helix destabilization effects on tRNA alone (44)(45)(46)(47). Similar observations have been made in RSV and MoMuLV, where nucleocapsid protein promotes the annealing of tRNA Trp and tRNA Pro to the PBSs of the RSV and MoMuLV genomic RNA, respectively (48).…”

Section: Viral and Cellular Proteins Regulating Trna Lys3 /Genomic Rnmentioning

confidence: 53%

“…This has been shown both in vitro (47,51,52) and in vivo (53). In the latter report, it was also shown that nucleocapsid protein mutations in Pr55 gag had a much greater effect on inhibiting tRNA Lys3 annealing than nucleocapsid protein mutations in Pr160 gag-pol , indicating an important role for Pr55 gag in tRNA Lys3 annealing.…”

Section: Viral and Cellular Proteins Regulating Trna Lys3 /Genomic Rnmentioning

confidence: 65%

tRNALys3: The Primer tRNA for Reverse Transcription in HIV‐1

Kleiman

2002

IUBMB Life

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“…However, both NC proteins have a high charge density, and it is known that this property is sufficient for facilitating nucleic acid rearrangements involving small numbers of base pairs, such as those involved in annealing tRNA to the primer-binding site of the genomic RNA (27). In contrast, a highly charged peptide cannot effectively facilitate minus-strand transfer in HIV-1 (9).…”

Section: Resultsmentioning

confidence: 99%

Specific zinc-finger architecture required for HIV-1 nucleocapsid protein's nucleic acid chaperone function

Williams

Gorelick²,

Musier‐Forsyth³

2002

Proc. Natl. Acad. Sci. U.S.A.

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108

144

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The nucleocapsid protein (NC) of HIV type 1 (HIV-1) is a nucleic acid chaperone that facilitates the rearrangement of nucleic acid secondary structure during reverse transcription. HIV-1 NC contains two CCHC-type zinc binding domains. Here, we use optical tweezers to stretch single -DNA molecules through the helix-to-coil transition in the presence of wild-type and several mutant forms of HIV-1 NC with altered zinc-finger domains. Although all forms of NC lowered the cooperativity of the DNA helix-coil transition, subtle changes in the zinc-finger structures reduced NC's effect on the transition. The change in cooperativity of the DNA helix-coil transition correlates strongly with in vitro nucleic acid chaperone activity measurements and in vivo HIV-1 replication studies using the same NC mutants. Moreover, Moloney murine leukemia virus NC, which contains a single zinc finger, had little effect on transition cooperativity. These results suggest that a specific two-zincfinger architecture is required to destabilize nucleic acids for optimal chaperone activity during reverse transcription in complex retroviruses such as HIV-1.

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HIV-1 nucleocapsid protein zinc finger structures induce tRNALys,3 structural changes but are not critical for primer/template annealing11Edited by M. F. Summers

Cited by 84 publications

References 73 publications

Differing Roles of the N- and C-terminal Zinc Fingers in Human Immunodeficiency Virus Nucleocapsid Protein-enhanced Nucleic Acid Annealing

Differing Roles of the N- and C-terminal Zinc Fingers in Human Immunodeficiency Virus Nucleocapsid Protein-enhanced Nucleic Acid Annealing

tRNALys3: The Primer tRNA for Reverse Transcription in HIV‐1

Specific zinc-finger architecture required for HIV-1 nucleocapsid protein's nucleic acid chaperone function

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