Match!

Ordered aggregation of ribonucleic acids by the human immunodeficiency virus type 1 nucleocapsid protein

Published on Mar 1, 1997in Biopolymers1.84
· DOI :10.1002/(SICI)1097-0282(199703)41:3<301::AID-BIP5>3.0.CO;2-W
Stoyl P. Stoylov11
Estimated H-index: 11
(CNRS: Centre national de la recherche scientifique),
Constance Vuilleumier7
Estimated H-index: 7
(CNRS: Centre national de la recherche scientifique)
+ 4 AuthorsYves Mély52
Estimated H-index: 52
(CNRS: Centre national de la recherche scientifique)
Cite
Abstract
The nucleocapsid protein NCp7, which is the major genomic RNA binding protein of human immunodeficiency virus type 1, plays an important role in several key steps of the viral life cycle. Many of the NCp7 activities, notably the nucleic acid annealing and the genomic RNA wrapping ones, are thought to be linked to a nonspecific binding of NCp7 to its nucleic acid targets. The mechanism of these activities is still debated but several clues are in favor of an intermediate aggregation of nucleic acids by NCp7. To check and characterize the nucleic acid aggregating properties of NCp7, we investigated the interaction of NCp7 with the model RNA homopolymer, polyA, by quasielastic light scattering and optical density measurements. The ordered growth of monodisperse large particles independently of the nucleic acid size and the almost complete covering of polyA by NCp7 strongly suggested an ordered aggregation mechanism. The aggregate kinetics of growth in the optimum protein concentration range (≥2 μM) were governed by a so-called Ostwald ripening mechanism limited by transfer of NCp7-covered polyA complexes from small to large aggregates. The aggregation process was strongly dependent on both Na+ and Mg2+ concentrations, the optimum concentrations being in the physiological range. Similar conclusions held true when polyA was replaced by 16S + 23S ribosomal RNA, suggesting that the NCp7 aggregating properties were only poorly dependent on the nucleic acid sequence and structure. Finally, as in the NCp7 annealing activities, the basic regions of NCp7, but not the zinc fingers, were found critical in nucleic acid aggregation. Taken together, our data indicate that NCp7 is a highly efficient nucleic acid aggregating agent and strengthen the hypothesis that aggregation may constitute a transient step in various NCp7 functions. © 1997 John Wiley & Sons, Inc.
  • References (45)
  • Citations (91)
Cite
References45
Newest
Published on Dec 31, 2008in Protein Science2.42
Fadia Dib-Hajj1
Estimated H-index: 1
(A&M: Texas A&M University),
Raza Khan7
Estimated H-index: 7
(A&M: Texas A&M University),
David P. Giedroc44
Estimated H-index: 44
(A&M: Texas A&M University)
The nucleocapsid protein (NC) is the major genomic RNA binding protein that plays integral roles in the structure and replication of all animal retroviruses. In this report, select biochemical properties of recombinant Mason-Pfizer monkey virus (MPMV) and HIV-1 NCs are compared. Evidence is presented that two types of saturated Zn2 NC-polynucleotide complexes can be formed under conditions of low [NaCl] that differ in apparent site-size (n = 8 vs. n = 14). The formation of one or the other compl...
Published on Oct 1, 1995in Journal of Molecular Biology5.07
Valérie Tanchou1
Estimated H-index: 1
('ENS Paris': École Normale Supérieure),
Caroline Gabus3
Estimated H-index: 3
('ENS Paris': École Normale Supérieure)
+ 1 AuthorsJean Luc Darlix2
Estimated H-index: 2
('ENS Paris': École Normale Supérieure)
Abstract HIV genomic RNA resides within the nucleocapsid, in the interior of the virus, which serves to protect the RNA against nuclease degradation and to promote its reverse transcription. To investigate the role of nucleocapsid protein (NCp7) in the stability and replication of genomic RNA within the nucleocapsid, we used NCp7, reverse transcriptase (RT) and RNAs representing the 5′ and 3′ regions of the genome to reconstitute functional HIV-1 nucleocapsids. The nucleoprotein complexes genera...
Published on Apr 1, 1995in Journal of Virology4.32
Jared L. Clever10
Estimated H-index: 10
,
C Sassetti1
Estimated H-index: 1
,
Tristram G. Parslow24
Estimated H-index: 24
The selective encapsidation of retroviral RNA requires sequences in the Gag protein, as well as a cis-acting RNA packaging signal (psi site) near the 5' end of the genomic transcript. Gag protein of human immunodeficiency virus type 1 (HIV-1) has recently been found to bind specifically to the HIV-1 psi element in vitro. Here we report studies aimed at mapping features within the genetically defined psi locus that are required for binding of HIV-1 Gag or of its processed nucleocapsid derivative....
Published on Jan 27, 1995in Journal of Biological Chemistry
Yves Mély52
Estimated H-index: 52
(CNRS: Centre national de la recherche scientifique),
Hugues de Rocquigny23
Estimated H-index: 23
(French Institute of Health and Medical Research)
+ 4 AuthorsDominique Gerard29
Estimated H-index: 29
(CNRS: Centre national de la recherche scientifique)
Abstract The nucleocapsid protein NCp7 of human immunodeficiency virus, type 1, is a key component in the viral life cycle. Since, the first common step of all reported NCp7 activities corresponds to a nucleic acid-binding step, the NCp7 binding parameters to the natural primer tRNA3 were investigated. Using NCp7 intrinsic fluorescence, we found that (i) in 0.1 M NaCl, NCp7 bound noncooperatively to tRNA3 with a K = 3.2 × 106M association constant and a n = 6 binding site size, (ii) four ionic i...
Published on Jan 1, 1995in Nucleic Acids Research11.15
Mary Lapadat-Tapolsky2
Estimated H-index: 2
,
Christine Pernelle1
Estimated H-index: 1
+ 1 AuthorsJean-Luc Darlix51
Estimated H-index: 51
Abstract Retroviral nucleocapsid (NC) protein is an integral part of the virion nucleocapsid where it is in tight association with genomic RNA and the tRNA primer. NC protein is necessary for the dimerization and encapsidation of genomic RNA, the annealing of the tRNA primer to the primer binding site (PBS) and the initial strand transfer event. Due to the general nature of NC protein-promoted annealing, its use to improve nucleic acid interactions in various reactions can be envisioned. Paramet...
Published on Oct 1, 1994in Journal of Virology4.32
Nijing Sheng1
Estimated H-index: 1
,
S. Erickson-Viitanen1
Estimated H-index: 1
Abstract The human immunodeficiency virus (HIV) gag polyprotein is processed by the viral protease to yield the structural proteins of the virus. One of these structural proteins, p15, and its protease cleavage products, p7 and p6, are believed to be responsible for the viral RNA binding which is prerequisite for assembly of infectious virions. To better understand potential interactions between viral RNA, p15, and the HIV protease, we have synthesized p15 in an in vitro system and studied its p...
Published on Sep 9, 1994in Journal of Biological Chemistry
Raza Khan7
Estimated H-index: 7
(A&M: Texas A&M University),
David P. Giedroc44
Estimated H-index: 44
(A&M: Texas A&M University)
Abstract The nucleocapsid protein (NC) of all animal retroviruses is the major structural protein of the core ribonucleoprotein complex, bound to genomic RNA in mature virions. In a previous report, we showed that recombinant NC protein from HIV-1, a 71-amino-acid protein (NC71), is apparently able to form two types of protein-nucleic acid complexes under low [NaCl], pH 8.3 and 25 degrees C. These appeared to differ in occluded apparent site size, napp, forming n = 8 and n = 14 complexes on poly...
Published on Sep 1, 1994in Journal of Virology4.32
Z Tsuchihashi1
Estimated H-index: 1
,
Patrick O. Brown145
Estimated H-index: 145
Abstract Nucleocapsid protein (NC) of human immunodeficiency virus type 1 (HIV-1) was expressed in Escherichia coli and purified. The protein displayed a variety of activities on DNA structure, all reflecting an ability to promote transition between double-helical and single-stranded conformations. We found that, in addition to its previously described ability to accelerate renaturation of complementary DNA strands, the HIV-1 NC protein could substantially lower the melting temperature of duplex...
Published on Jul 1, 1994in Virology2.66
Robert D. Berkowitz1
Estimated H-index: 1
(HHMI: Howard Hughes Medical Institute),
Stephen P. Goff71
Estimated H-index: 71
(HHMI: Howard Hughes Medical Institute)
Abstract We previously used RNA gel mobility shift assays to demonstrate specific binding of the HIV-1 gag precursor polyprotein and nucleocapsid (NC) protein to HIV-1 RNA and to map the binding elements in each species by mutagenesis. Here we report finer mapping of binding elements in the HIV-1 genomic RNA and NC protein, performed by analyzing the binding behavior of fragments of each species in the gel shift assay. With regard to the RNA, the strongest binding activity resided in a 120-nucle...
Cited By91
Newest
Published on Feb 1, 2018in European Journal of Medicinal Chemistry4.83
Elia Gamba1
Estimated H-index: 1
(UNIPD: University of Padua),
Mattia Mori14
Estimated H-index: 14
(UNISI: University of Siena)
+ 7 AuthorsMaurizio Botta43
Estimated H-index: 43
(UNISI: University of Siena)
Abstract In this report, we present a new benzoxazole derivative endowed with inhibitory activity against the HIV-1 nucleocapsid protein (NC). NC is a 55-residue basic protein with nucleic acid chaperone properties, which has emerged as a novel and potential pharmacological target against HIV-1. In the pursuit of novel NC-inhibitor chemotypes, we performed virtual screening and in vitro biological evaluation of a large library of chemical entities. We found that compounds sharing a benzoxazolino...
Published on Feb 1, 2018in RNA3.95
Shringar Rao3
Estimated H-index: 3
,
Alessandro Cinti5
Estimated H-index: 5
+ 3 AuthorsAndrew J. Mouland29
Estimated H-index: 29
Published on Feb 23, 2017in Journal of Physical Chemistry B2.92
Avisek Ghose3
Estimated H-index: 3
,
Oleg V. Maltsev14
Estimated H-index: 14
+ 5 AuthorsPascal Didier18
Estimated H-index: 18
In this work, we used firefly oxyluciferin (OxyLH2) and its polarity-dependent fluorescence mechanism as a sensitive tool to monitor biomolecular interactions. The chromophores, OxyLH2, and its two analogues, 4-MeOxyLH and 4,6′-DMeOxyL, were modified trough carboxylic functionalization and then coupled to the N-terminus part of Tat and NCp7 peptides of human immunodeficiency virus type-1 (HIV-1). The photophysical properties of the labeled peptides were studied in live cells as well as in comple...
Published on Dec 1, 2016in Retrovirology3.74
Klara Post5
Estimated H-index: 5
(NIH: National Institutes of Health),
Erik D. Olson5
Estimated H-index: 5
(OSU: Ohio State University)
+ 5 AuthorsJudith G. Levin37
Estimated H-index: 37
(NIH: National Institutes of Health)
Background The nucleocapsid (NC) domain of HIV-1 Gag is responsible for specific recognition and packaging of genomic RNA (gRNA) into new viral particles. This occurs through specific interactions between the Gag NC domain and the Psi packaging signal in gRNA. In addition to this critical function, NC proteins are also nucleic acid (NA) chaperone proteins that facilitate NA rearrangements during reverse transcription. Although the interaction with Psi and chaperone activity of HIV-1 NC have been...
Published on Dec 1, 2016in Retrovirology3.74
Katarzyna Pachulska-Wieczorek7
Estimated H-index: 7
(PAN: Polish Academy of Sciences),
Leszek Błaszczyk3
Estimated H-index: 3
(PUT: Poznań University of Technology)
+ 2 AuthorsKatarzyna J. Purzycka12
Estimated H-index: 12
(PAN: Polish Academy of Sciences)
Background The Gag polyprotein is a multifunctional regulator of retroviral replication and major structural component of immature virions. The nucleic acid chaperone (NAC) activity is considered necessary to retroviral Gag functions, but so far, NAC activity has only been confirmed for HIV-1 and RSV Gag polyproteins. The nucleocapsid (NC) domain of Gag is proposed to be crucial for interactions with nucleic acids and NAC activity. The major function of matrix (MA) domain is targeting and bindin...
Published on Sep 3, 2015in Nucleic Acids Research11.15
Yuri Nishida3
Estimated H-index: 3
(UGA: University of Georgia),
Katarzyna Pachulska-Wieczorek7
Estimated H-index: 7
(PAN: Polish Academy of Sciences)
+ 4 AuthorsKatarzyna J. Purzycka12
Estimated H-index: 12
(PAN: Polish Academy of Sciences)
Ty1 Gag comprises the capsid of virus-like particles and provides nucleic acid chaperone (NAC) functions during retrotransposition in budding yeast. A subgenomic Ty1 mRNA encodes a truncated Gag protein (p22) that is cleaved by Ty1 protease to form p18. p22/p18 strongly inhibits transposition and can be considered an element-encoded restriction factor. Here, we show that only p22 and its short derivatives restrict Ty1 mobility whereas other regions of GAG inhibit mobility weakly if at all. Mutat...
Published on Mar 1, 2015in Biochemical and Biophysical Research Communications2.71
Dominic F. Qualley4
Estimated H-index: 4
(Berry College),
Victoria L. Sokolove1
Estimated H-index: 1
(Berry College),
James L. Ross3
Estimated H-index: 3
(Berry College)
Abstract Nucleocapsid proteins (NCs) direct the rearrangement of nucleic acids to form the most thermodynamically stable structure, and facilitate many steps throughout the life cycle of retroviruses. NCs bind strongly to nucleic acids (NAs) and promote NA aggregation by virtue of their cationic nature; they also destabilize the NA duplex via highly structured zinc-binding motifs. Thus, they are considered to be NA chaperones. While most retroviral NCs are structurally similar, differences are o...
Published on Jan 1, 2015in Current Topics in Microbiology and Immunology3.15
Mattia Mori14
Estimated H-index: 14
(UNISI: University of Siena),
Lesia Kovalenko2
Estimated H-index: 2
(UDS: University of Strasbourg)
+ 5 AuthorsYves Mély52
Estimated H-index: 52
(UDS: University of Strasbourg)
Open image in new window HIV reverse transcription and nucleocapsid. After the capsid has entered the cell, reverse transcriptase (A) creates a DNA copy (green) of the HIV RNA genome (yellow), using a cellular transfer RNA (B) as a primer. HIV nucleocapsid protein (C) acts as a chaperone to unfold the RNA secondary structure. The ribonuclease activity of RT removes the viral RNA after the DNA strand is created. Interaction of HIV Vif (D) with cellular APOBEC (E) is also shown
Published on Dec 1, 2014in Retrovirology3.74
Katarzyna Pachulska-Wieczorek7
Estimated H-index: 7
(PAN: Polish Academy of Sciences),
Agnieszka K Stefaniak1
Estimated H-index: 1
(PAN: Polish Academy of Sciences),
Katarzyna J. Purzycka12
Estimated H-index: 12
(PAN: Polish Academy of Sciences)
Background The nucleocapsid domain of Gag and mature nucleocapsid protein (NC) act as nucleic acid chaperones and facilitate folding of nucleic acids at critical steps of retroviral replication cycle. The basic N-terminus of HIV-1 NC protein was shown most important for the chaperone activity. The HIV-2 NC (NCp8) and HIV-1 NC (NCp7) proteins possess two highly conserved zinc fingers, flanked by basic residues. However, the NCp8 N-terminal domain is significantly shorter and contains less positiv...
View next paperNucleic-acid-chaperone activity of retroviral nucleocapsid proteins: significance for viral replication.