Cloning and characterization of a cell surface receptor for xenotropic and polytropic murine leukemia viruses - PubMed (original) (raw)

Cloning and characterization of a cell surface receptor for xenotropic and polytropic murine leukemia viruses

C S Tailor et al. Proc Natl Acad Sci U S A. 1999.

Abstract

Xenotropic and polytropic murine leukemia viruses (X-MLVs and P-MLVs) cross-interfere to various extents in non-mouse species and in wild Asian mice, suggesting that they might use a common receptor for infection. Consistent with this hypothesis, the susceptibility of some wild mice to X-MLVs has been mapped to the P-MLV receptor locus at the distal end of mouse chromosome 1. In this study, we report the isolation and characterization of a cDNA for the human X-MLV cell surface receptor (X-receptor) by using a human T lymphocyte cDNA library in a retroviral vector. The predicted X-receptor contains 696 amino acids with multiple hydrophobic potential membrane-spanning sequences and with weak homologies to the yeast proteins SYG1, of unknown function, and PHO81, which has been implicated in a system that regulates transport of inorganic phosphate. Expression of the X-receptor in Chinese hamster ovary cells, which are substantially resistant to P-MLVs and to X-MLVs, made them susceptible to both of these virus groups. The mouse homologue of the X-receptor was mapped by hybridization to the distal end of chromosome 1 at the same position as the P-MLV receptor gene Rmc1. These results strongly support the hypothesis that a common gene encodes the receptors for X-MLVs and P-MLVs, with the human X-receptor preferentially mediating X-MLV infections and the homologous protein of inbred mice mediating only P-MLV infections. We propose that X-MLVs and P-MLVs comprise a single family of retroviruses that have coevolved in response to diversification in X-receptor genes of the host.

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Figures

Figure 1

Figure 1

Susceptibility of NIH 3T3 cells expressing the X3 cDNA to X-MLV pseudotype infection. Target cells were analyzed for their susceptibility to lacZ(X-MLV) pseudotype. (A) Infection of the puromycin-resistant NIH 3T3 cellular clone X3, previously transduced with the retroviral cDNA library vector and then with puro(X-MLV) pseudotype (see Materials and Methods). (B) Infection of NIH 3T3 cells. (C) Infection of NIH 3T3 cells transfected with pcDNA3.1-X3 (X3 cDNA cloned into the pCDNA3.1 mammalian expression vector). (D) Infection of NIH 3T3 cells transfected with only pCDNA3.1 vector.

Figure 2

Figure 2

The amino acid sequence and hydrophobicity plot of X-receptor and sequence comparison to the yeast SYG1 protein. (A) The X-receptor is a large protein of 696 amino acids. Potential membrane-spanning domains (underlined) were identified by using the Kyte and Doolittle algorithm (35). Potential N-linked glycosylation sites (NXS/T) are shown by asterisks. (B) Hydrophobicity plots were generated by using the Kyte and Doolittle algorithm. X-receptor and SYG1 contained at least eight hydrophobic potential membrane spanning sequences, as indicated. The transmembrane region labeled 5 could possibly contain an additional membrane-spanning sequence. (C) Alignment of highly related regions between SYG1 and X-receptor (X) determined by

blast

comparisons. The consensus sequence is shown between the X-receptor and SYG1 sequence, where + indicates related amino acids.

Figure 3

Figure 3

Northern blot analysis of Poly(A)+ RNA from various human tissues. The multiple-tissue Northern blots from human were probed with [32P]-labeled X3 cDNA. FL, fetal liver; BM, bone marrow; PBL, peripheral blood lymphocytes; T, thymus; LN, lymph node; S, spleen; P, pancreas; K, kidney; SM, skeletal muscle; L, liver; Lg, lung; Pl, placenta; B, brain; H, heart.

Figure 4

Figure 4

Map location of the xenotropic/polytropic receptor gene on mouse Chr1. The map represents a 10-centimorgan segment of the distal end of Chr1. The entire Chr1 has a length of ≈107 centimorgans. To the right of the map are recombination fractions between adjacent loci, with the first fraction from the M. musculus musculus crosses and the second from M. spretus crosses. The numbers in parentheses represent recombinational distances ± standard errors. Marker loci were typed as previously described in this cross [C. A. Kozak and C. E. Buckler, Mouse Genome Database, Release 3.2, Mouse Genome Informatics, The Jackson Laboratory (

http://www.informatics.jax.org/crossdata

); ref. 38). The M. spretus crosses were not typed for Lamc1. Gene order was established by minimizing the number of recombinants. One single locus double recombinant involving Cchl2a was identified in the M. spretus crosses that is not included in the figure.

Comment in

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