PubMed Health⌕ Search

PubMed · 11597756

Hypervariable genes in Grapevine leafroll associated virus 1.

Abstract

Analysis of nucleotide sequences of 10 open reading frames from the Grapevine leafroll associated virus 1 (GLRaV-1), a tentative member of the genus Closterovirus, revealed the presence of an unusually high degree of sequence variation in ORFs 3, 6 and 7 encoding a homologue of heat shock protein 70 and two diverged copies of the coat protein (CPd1 and CPd2), respectively. Overall, 75 clones corresponding to ORFs 3, 6 and 7 were sequenced and 1916 nucleotide changes were recorded relative to the published sequence. Surprisingly, none of the changes resulted in a frame shift or stop codon and there was a trend for the conservation of amino acids or change to amino acids having similar physiochemical properties. The CPd2 gene was particularly variable with a mutation seen in 60% of the nucleotide positions in one or more of the 1.1-kb cDNA clones sequenced. These observations suggest that GLRaV-1 may exist in the form of a heterogeneous population, possibly resulting from the lack of selective pressure and from mixing of virus strains due to viticulture practices of vegetative propagation and grafting over the centuries.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

A Little, C F Fazeli, M A Rezaian. 2001-11-28. Hypervariable genes in Grapevine leafroll associated virus 1.. https://doi.org/10.1016/s0168-1702(01)00343-4

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

A directed evolution approach to select for novel Adeno-associated virus capsids on an HIV-1 producer T cell line.

A directed evolution approach was used to select for Adeno-associated virus (AAV) capsids that would exhibit more tropism toward an HIV-1 producer T cell line with the long-term goal of developing improved gene transfer vectors. A library of AAV variants was used to infect H9 T cells previously infected or uninfected by HIV-1 followed by AAV amplification with wild-type adenovirus. Six rounds of biological selection were performed, including negative selection and diversification after round three. The H9 T cells were successfully infected with all three wild-type viruses (AAV, adenovirus, and HIV-1). Four AAV cap mutants best representing the small number of variants emerging after six rounds of selection were chosen for further study. These mutant capsids were used to package an AAV vector and subsequently used to infect H9 cells that were previously infected or uninfected by HIV-1. A quantitative polymerase chain reaction assay was performed to measure cell-associated AAV genomes. Two of the four cap mutants showed a significant increase in the amount of cell-associated genomes as compared to wild-type AAV2. This study shows that directed evolution can be performed successfully to select for mutants with improved tropism for a T cell line in the presence of HIV-1.

Capsid↗

Engineering adeno-associated viruses for clinical gene therapy.

Clinical gene therapy has been increasingly successful owing both to an enhanced molecular understanding of human disease and to progressively improving gene delivery technologies. Among these technologies, delivery vectors based on adeno-associated viruses (AAVs) have emerged as safe and effective and, in one recent case, have led to regulatory approval. Although shortcomings in viral vector properties will render extension of such successes to many other human diseases challenging, new approaches to engineer and improve AAV vectors and their genetic cargo are increasingly helping to overcome these barriers.

Capsid↗

The pH dependence of HIV-1 capsid assembly and its interaction with cyclophilin A.

Immature HIV-1 virions have spherical cores which become conical due to cleavage of the capsid domain of Gag. Here, we have used an immature form of capsid and show by electron microscopy, atomic force microscopy and single angle light scattering that it aggregates to spherical cores resembling immature virions at high ionic strengths and at pH values above 6. Dynamic angle light scattering of the dissociated protein shows structural changes that promote oligomerization above pH 6. We then examined the role of the required host protein cyclophilin A on assembly. Cyclophilin A is incorporated into virions at a 1:10 cyclophilin A/capsid ratio. We find that although cyclophilin A does not affect the oligomerization rate or stability of immature capsid cores, it does bind strongly to immature capsid at physiological stoichiometry above pH 6. This association serves as an entry route of cyclophilin A into HIV-1 virions.

Capsid↗