Possible roles of nucleocapsid protein of MoMuLV in the specificity of proviral DNA synthesis and in the genetic variability of the virus

Autor: Jean-Luc Darlix, Damien Ficheux, Jean-Baptiste Rascle
Přispěvatelé: Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Deleage, Gilbert
Rok vydání: 1998
Předmět:
Zdroj: Journal of Molecular Biology
Journal of Molecular Biology, Elsevier, 1998, 280, pp.215-225
ISSN: 0022-2836
1089-8638
DOI: 10.1006/jmbi.1998.1873
Popis: International audience; Retroviral nucleocapsid (NC) protein, in addition to its structural roles in the virion core, is involved in the early and late phases of the viral replication cycle. To further characterise the role of NC protein of MoMuLV (NCp10) in the replication of the viral genome, the influence of NCp10 on self-primed versus primer-specific reverse transcription has been analysed in vitro. The results show that NCp10 can enhance the specificity of proviral DNA synthesis by inhibiting self-primed cDNA synthesis while promoting primer-specific DNA synthesis within active NCp10-RNA nucleoprotein complexes. Retroviruses are known to show a high degree of variability and this prompted us to examine the possible implication of NCp10 in the genetic variability of MoMuLV. The ability of reverse transcriptase (RT) to extend different mutated primers using an RNA or a DNA template has been investigated in the presence or in the absence of NCp10. NCp10 was found to have different effects on RT depending on the nature of the template: an enhancement at the elongation level of mutated primers using RNA as template versus a slight inhibition using DNA as template. These observations suggest that NCp10 could be implicated in the genetic variability of MoMuLV by allowing nucleotide misincorporation principally during minus strand DNA synthesis.Retroviral nucleocapsid (NC) protein, in addition to its structural roles in the virion core, is involved in the early and late phases of the viral replication cycle. To further characterise the role of NC protein of MoMuLV (NCp10) in the replication of the viral genome, the influence of NCp10 on self-primed versus primer-specific reverse transcription has been analysed in vitro. The results show that NCp10 can enhance the specificity of proviral DNA synthesis by inhibiting self-primed cDNA synthesis while promoting primer-specific DNA synthesis within active NCp10-RNA nucleoprotein complexes. Retroviruses are known to show a high degree of variability and this prompted us to examine the possible implication of NCp10 in the genetic variability of MoMuLV. The ability of reverse transcriptase (RT) to extend different mutated primers using an RNA or a DNA template has been investigated in the presence or in the absence of NCp10. NCp10 was found to have different effects on RT depending on the nature of the template: an enhancement at the elongation level of mutated primers using RNA as template versus a slight inhibition using DNA as template. These observations suggest that NCp10 could be implicated in the genetic variability of MoMuLV by allowing nucleotide misincorporation principally during minus strand DNA synthesis.
Databáze: OpenAIRE