Interactions in vivo between the Vif protein of HIV-1 and the precursor (Pr55GAG) of the virion nucleocapsid proteins

The abnormality of viral core structure seen in vif-defective HIV-1 grown in PBMCs has suggested a role for Vif in viral morphogenesis. Using an in vivo mammalian two-hybrid assay, the interaction between Vif and the precursor (Pr55GAG) of the virion nucleocapsid proteins has been analysed. This revealed the amino-terminal (aa 1–22) and central (aa 70–100) regions of Vif to be essential for its interaction with Pr55GAG, but deletion of the carboxy-terminal (aa 158–192) region of the protein had only a minor effect on its interaction. Initial deletion studies carried out on Pr55GAG showed that a 35-amino-acid region of the protein bridging the MA(p17)–CA(p24) junction was essential for its ability to interact with Vif. Site-directed mutagenesis of a conserved tryptophan (Trp21) near the amino terminus of Vif showed it to be important for the interaction with Pr55GAG. By contrast, mutagenesis of the highly conserved YLAL residues forming part of the BC-box motif, shown to be important in Vif promoting degradation of APOBEC3G/3F, had little or no effect on the Vif–Pr55GAG interaction

Polymorphisms in Gag spacer peptide 1 confer varying levels of resistance to the HIV- 1maturation inhibitor bevirimat

Background: The maturation inhibitor bevirimat (BVM) potently inhibits human immunodeficiency virus type 1 (HIV-1) replication by blocking capsid-spacer peptide 1 (CA-SP1) cleavage. Recent clinical trials demonstrated that a significant proportion of HIV-1-infected patients do not respond to BVM. A patient’s failure to respond correlated with baseline polymorphisms at SP1 residues 6-8. Results: In this study, we demonstrate that varying levels of BVM resistance are associated with point mutations at these residues. BVM susceptibility was maintained by SP1-Q6A, -Q6H and -T8A mutations. However, an SP1-V7A mutation conferred high-level BVM resistance and SP1-V7M and T8Δ mutations conferred intermediate levels of BVM resistance. Conclusions: Future exploitation of the CA-SP1 cleavage site as an antiretroviral drug target will need to overcome the baseline variability in the SP1 region of Gag.Publisher PDFPeer reviewe

Thymosin beta arg10, a major variant of thymosin beta 10 in rabbit tissues.

Two homologous peptides, designated thymosin beta 4 and thymosin beta 10, respectively, have been shown to be widely distributed in mammalian cells and tissues (S. Erickson-Viitanen, S. Ruggieri, P. Natalini, and B.L. Horecker (1983) Arch. Biochem. Biophys. 221, 570-576; S. Erickson-Viitanen, S. Ruggieri, P. Natalini, and B.L. Horecker, (1983) Arch. Biochem. Biophys. 225, 407-413). In the rabbit, thymosin beta 4 is replaced by a variant, thymosin beta ala4, that contains alanine in place of serine at the blocked NH2-terminus. It is reported that in rabbit tissues thymosin beta 10 is also replaced by a variant, designated thymosin beta arg10, that contains an additional amino acid, arginine, inserted following lysine-38. The rabbit tissues analyzed also differ from those of other mammals in the relative quantities of thymosin beta ala4 and beta arg10, which are nearly equal, compared to tissues from other mammals where the quantities of thymosin beta 10 are only one-third to one-tenth those of thymosin beta 4

Thymosin beta 10, a new analog of thymosin beta 4 in mammalian tissues.

A new analog of thymosin beta 4 has been isolated from tissues of several mammalian species. This peptide, designated thymosin beta 10, is composed of 42 amino acid residues and shows 75% sequence homology with thymosin beta 4. It occurs together with thymoxin beta 4 in a variety of tissues including spleen, liver, and thymus and also in several cultured cell lines. In the spleen of rat, mouse, cat, and man, the new peptide accounts for approximately 0.02% by weight of the total protein. In the calf it is replaced by another homologous peptide, designated thymosin beta 9, whose structure has been reported

Distribution of Thymosin beta 4 in Vertebrate Classes

A peptide containing 43 amino acid residues, rich in glutamic acid and lysine, was originally isolated from calf thymus and designated thymosin beta 4 [T.L.K. Low, S. -K. Hu, and A. L. Goldstein (1981) Proc. Nat. Acad. Sci. USA 78, 1162-1166]. However, thymosin beta 4 was also shown to be present in other tissues of rats and mice, with highest concentrations in spleen and peritoneal macrophages [E. Hannappel, G.-J. Xu, J. Morgan, J. Hempstead, and B.L. Horecker (1982), Proc. Nat. Acad. Sci. USA 79, 2172-2175]. We have now identified the same peptide in tissues of other mammalian species and other vertebrate classes, including birds and amphibia. Exceptions are the rabbit and bony fish, where thymosin beta 4 is replaced by different peptides, similar in size and in amino acid sequence. None of these peptides was detected in several invertebrates or in the protozoan, Tetrahymena pyriformis. In subcellular fractionation of rat spleen, thymosin beta 4 was recovered in the cytosol