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MaTu is an agent, believed to be derived from a human mammary carcinoma, which displayed several extraordinary properties. These were: RIP and PAGE revealed in MaTu-infected cells only a single protein band of Mr 58 k, the gp 58. This gp 58 was immunoprecipitated by antibodies present in some human sera as well as in some sera of rabbits, sheep, and cattle. MaTu had an extremely restricted host range: it was transmissible only to HeLa cells, but not to human embryo fibroblasts, to three human tumour cell lines (T 47 D, T 24, and HMB 2) or to monkey Vero and rabbit SIRC cells. A retrovirus with a broad host range, used as a helper (X-MLV) enabled the transmission of MaTu to human fibroblasts, but not to Vero or SIRC, which are also permissive for X-MLV. These observations, together with our previous reports, support the view that MaTu might either be a novel type of defective virus, or even a non-viral autonomous genetic element.

We have identified the major immunogenic structural proteins of the human foamy virus (HFV), a distinct member of the foamy virus subfamily of Retroviridae. Radiolabelled viral proteins were immunoprecipitated from HFV-infected cells by foamy virus antisera of human and non-human primate origin. Precipitated viral proteins were in the range of 31K to 170K. Labelling of proteins with [14C]glucosamine or with [35S]methionine in the presence of tunicamycin, as well as endo-beta-N-acetylglycosaminidase H and F treatment of [35S]methionine-labelled proteins, revealed three viral glycoproteins of approximately 170K, 130K and 47K, most likely representing the env gene-encoded precursor, the surface glycoprotein and the transmembrane protein of HFV, respectively.

An epithelial cell line (RC-4B/C) was established from a pituitary adenoma obtained from a 3-yr-old (ACI/fMai X F344/fMai)F1 male rat. Before Year 5 in vitro, RC-4B/C cells could not be viably recovered from cryogenic storage. Recovery of viable cells from cryogenic storage in Year 5 was associated with a more transformed phenotype, including the appearance of endogenous C-type rat retroviral particles. The ultrastructural appearance of the cells was similar to that of differentiated anterior pituitary cells; the cultured cells contained numerous, electron dense, secretory granules, Golgi complexes, and extended arrays of rough endoplasmic reticulum. Immunocytochemical study showed that all cell types present in the rat anterior pituitary gland were present in the cell line. The percentage of luteinizing hormone beta (LH beta) cells in the cell line was higher (19.9%) and that of growth hormone cells was lower (12.2%) than in normal male rat pituitary, whereas the cell line contained a comparable percentage of follicle stimulating hormone beta (FSH beta), prolactin (PRL), ACTH, and thyrotropin beta cells. Radioimmunoassay data demonstrated the PRL content of the cells was comparable to that of normal male rat pituitary gland, whereas the content of LH and FSH was 70- and 800-fold lower, respectively. Assay of specific receptor sites for gonadotropin releasing hormone (GnRH) using Scatchard plots of the data established the RC-4B/C cells contained GnRH receptor sites of the same affinity as in the pituitary gland, but of twofold lower capacity. These data suggest the RC-4B/C cell line warrants further study as a model for the induction and maintenance of the gonadotropic function of the pituitary gland.

We find that 12 of 14 specimens of normal human term placentas analyzed by one- or two-dimensional electrophoresis and immunoblotting contain a protein or polypeptide of approximately equal to 30,000 daltons that is antigenically cross-reactive with p30 core protein of the simian sarcoma-associated virus/gibbon ape leukemia virus primate retrovirus group and is physicochemically similar to reference murine and primate type C retrovirus p30s. This finding may lead to an understanding of endogenous type C retrovirus gene expression in humans.

The major internal protein of bovine leukemia virus (BLV p24) was isolated using ion exchange chromatography on phosphocellulose and gel filtration. The specificity of the BLV p24 isolated was checked by both the radioimmunoprecipitation (RIP) and the competitive radioimmunoassay (RIA). No cross reactivity between BLV p25 and the mammalian viruses of type C and D and the retrovirus isolated from human myeloma cells RPMI8226 [8, 17] was detected. The analysis of 429 leukemia-suspected bovine blood sera resulted in the detection of 165 (38.5%) positive and 264 (61.5%) negative blood sera. A correlation of the results of radioimmunoprecipitation reaction of major internal protein p24 and immunodiffusion test on the glycoprotein antigen of BLV was observed.

In contrast to the human cell line derived type D retrovirus PMFV, the Mason-Pfizer monkey virus (MPMV) does not suppress the mitogen response of normal human lymphocytes. Both viruses have been propagated on the same cell lines and purified by the same methods. MPMV did not contain a factor able to abolish PMFV-induced suppression of the mitogen response. Neither could MPMV suppress the mitogen response of lymphocytes from rhesus monkeys or baboons. PMFV however inhibited their reactivity.

A sensitive micro in situ fluorescent focus assay (mISFA), which utilizes cell monolayers fixed on a plastic surface, was developed for the detection of NZB xenotropic type C retroviruses grown in mammalian cell lines. The mISFA technique is an easier, more rapid and less expensive assay than other biological tests used to detect these viruses; it gives a more precise viral titer and is slightly more sensitive. This technique allows extension of host-range investigations to include any growing cell cultures and is applicable to various virus-host cell systems.

Cocultivation of uninfected and bovine leukemia virus-producing bat cells yielded, in addition to the unintegrated linear DNA duplex, DNA molecules that migrated as 4.4- and 4.8-kilobase-pair DNA fragments in gel electrophoresis. These DNA molecules were purified by acid-phenol extraction and cleaved with restriction endonucleases EcoRI, and HindIII, which have one recognition site each on the bovine leukemia virus proviral DNA. Such cleavage generated DNA molecules of approximately 10.0 and 9.4 kilobase pairs, thus indicating the existence of two species of covalently closed circular molecules of bovine leukemia virus proviral DNA.

Endogenous xenotropic cat type C virus (RD-114)- and infectious feline leukemia virus (FeLV)-specific gene expressions were measured in spontaneous sarcomas carcinomas, and nonmalignant cat tissues by molecular hybridization for virus-specific RNA and competition radio-immunoassays for the major internal protein (p30) of these two viruses. The results indicate that RD-114 gene expression in sarcomas and carcinomas at both RNA and p30 levels is significantly higher than histologically normal tissues from cats free of cancer. In contrast, the levels of FeLV viral RNA and p30 are fount to be low or undetectable in the majority of these tumored and normal tissues examined. Whereas variability in the amounts of RD-114 OR FeLV RNA and p30 expressed is found in tissues from different cats, their expression is fairly uniform in multiple malignant tissues of the same cat. The finding of widespread occurrence of elevated RD-114 gene expression in sarcomas and carcinomas is consistent with our similar observation with natural lymphomas of domestic cats and suggests that expression of certain functions of this endogenous virus may be etiologically involved in the development of many different spontaneous neoplasms of cats.

The RNA of a type D retrovirus (PMFV) isolated from and propagated in a human cell ine has been characterized. Like other retroviruses PMFV contains a 60 to 70 S RNA as determined by glycerol gradient velocity sedimentation with marker RNAs. When denatured by heat the high-molecular-weight RNA prepared from fresh virus yielded subunits of about 30 S. As shown by poly(U)-Sepharose chromatography the 60 to 70 S RNA as well as the 30 S subunits contain a poly (A) stretch of approximately 80 to 200 nucleotides.

The nucleotide sequences related to the 3'-terminal protion of retrovirus genomic RNA have been detected in the DNA of animals, including humans. The DNA complementary to the 400 to 700 nucleotides from the 3'-terminal end of retrovirus RNA (cDNA3'), which contains the enriched conserved region, was hybridized with DNA from a variety of animal cells. Under the conditions of annealing in 0.72 M NaCl at 67 degrees C and hydroxyapatite chromatography at 55 degrees C, 20 to 50% of the radioactivity of the cDNA3' prepared from two retroviruses, a murine Rauscher virus (RLV) and a baboon virus (M7), annealed with normal cellular DNA of animals, including human tissue. The thermal denaturation profile revealed considerable mismatching between the duplex of the cDNA3' and human DNA, cDNA3' of retroviruses is most homologous to cellular DNA of the host species of origin and is less homologous to cellular DNA of species that are distant in the phylogeny of the host species. The conservation and evolution of nucleotide sequences related to the 3' end of retrovirus genomes in animal DNAs, including humans, suggest that the sequences may have important functions.

The complete amino acid sequence of the nucleic acid-binding protein p12 of bovine leukemia virus (BLV) has been determined. Peptides were generated by enzymatic digestion and formic acid cleavage, purified by reversed-phase liquid chromatography and subjected to automated Edman degradation. BLV p12 is a proline-rich linear polypeptide composed of 69 amino acids with Mr 7558. A comparison of the p12 structure to that of the avian and murine type C retroviral nucleic acid-binding proteins shows significant homology only in the putative binding domain. This conserved region is duplicated BLV p12 as in the avian homolog.

The properties of murine oncornavirus produced by cells of spontaneous lymphosarcroma of CC57Br mice are described. In addition to the properties common for C-type RNA tumor viruses such as 60-70 S high molecular weight RNA, the presence of RNA-directed DNA polymerase and murine gs I (intraspecies) antigen and typical morphology in electron microscope, etc., the virus under study is characterized by instability of virions, unusual features of the DNA polymerase system and by the absence of demonstrable oncogenicity either in laboratory animals or in tissue cultures.

Seventeen placentas from normal Japanese women, obtained from 15 full term gestations and at 2 earlier gestational periods, were observed by electron microscopy for the presence of type C retrovirus-like particles. Immature free and budding particles were found in 16 placentas including those obtained pre-term. Free virus-like particles were seen in the intercellular spaces and within the lysosomes of syncytiotrophoblasts of the chorionic villi. Forms budding from syncytiotrophoblasts were found in 3 cases. Coincident data with the electron microscopic observations were obtained by immunohistological methods. Specific positive staining with antisera against heterogeneic whole murine leukaemia virus were seen within the cytoplasm of trophoblastic cells both in and along the basal lamina.

A technique originally described for the isolation of Friend leukaemia virus envelope polypeptides [1] yields equivalent structures from Moloney leukaemia. AKR and BALB/c xenotropic virus as well as feline leukaemia virus. The envelope polypeptides are obtained as micellar protein complexes, named rosettes. Rosettes of the five mammalian type-C viruses examined are indistinguishable by electron microscopy. Separation of these aggregates in polyacrylamide gel electrophoresis under nonreducing conditions reveals a glycoprotein of about 85000 d as their major component. Tryptic peptide analyses identify the viral origin of these polypeptides and emphasize strain specific differences in their primary structure.

Two murine sarcoma viruses, the Kirsten and the Harvey, were isolated by passage of mouse type C leukemia viruses through rats. These sarcoma viruses have genomes containing portions of their parental type C mouse leukemia virus genomes, in stable association with specific rat cellular sequences that we find to be quite likely not those of a rat type C leukemia virus. To determine if these murine sarcoma viruses provide a model relevant to the events occurring in spontaneous tumors, we have hybridized DNA and RNA prepared from rat tumors and normal rat tissues to [3H]DNA prepared from the Kirsten murine sarcoma virus. We have also hybridized these rat tissue nucleic acids to [3H]DNA prepared from a respresentative endogenous rat type C leukemia virus, the WFU (Wistar-Furth). Sarcoma-viral rat cellular sequences and endogenous rat leukemia viral sequences were detected in the DNA of both tumor and normal tissues, with no evidence of either gene amplification or additional sequences being present in tumor DNA. Sarcoma-viral rat cellular sequences and endogenous rat leukemia viral sequences were detected at elevated concentrations in the RNA of many rat tumors and in specific groups of normal tissues.

The results of molecular hybridization experiments have demonstrated that the RNA genome of RD-114 virus has extensive nucleotide sequence homology with the RNA genome of Crandell virus, an endogenous type C virus of cats, but only limited homology with the RNA genomes of feline sarcoma virus and feline leukemia virus. The genomic RNAs of RD-114 virus and Crandell virus also had identical sedimentation coefficients of 50S. A structural rearrangement of genomic RNA did not exist within released RD-114 virions, whereas a structural rearrangement of genomic RNA did occur within feline sarcoma virions and feline leukemia virions after release from virus-producing cells.

It was isolated and identified MR-Le virus from Wistar rats with myelogenous leukemia. The comparison of isolated in vivo and in vitro MR-Le virus with endogenous oncovirus RaLV in protein profile as well as results of molecular hybridization pointed out that RaLV did not participate in the induction of MR leukemia. The question about the role of MR-Le virus in the etiology of MR leukemia is opened now.

Three xenotropic retroviruses have been biologically cloned from cells cultured from the kidney of a 3-month-old NZB female mouse. They were obtained by first cocultivating the kidney cells for several weeks with mink, dog, and human cells and then cloning them by endpoint dilution. The cloned viruses differ in their infectivity and replicative ability in a variety of heterologous cell lines. The mink cell line-derived virus (X-NZB/K-Mlc) reaches titers in culture of over 10(8) infectious viruses/ml, and is produced in high titer within 24 hr after infection of mink lung cells. The human and dog cell-derived NZB viruses (X-NZB/K-Huc and X-NZB/K-Dgc) grow to lower titers and are similar in many respects. They differ in their relative ability to replicate in dog and human cells and to transform mink S+L- cells. Peptide mapping studies indicate that the X-NZB/K-Mlc virus has a unique p15(E) protein which distinguishes it from the other two cloned NZB viruses. These results lend further support to the observation that several types of xenotropic virus are present in a mouse strain and that more than one virus can be expressed by one organ of a particular mouse.

Clonal isolates of an early passage stock of woolly monkey sarcoma virus (WSV) have been shown to code for different numbers of woolly monkey helper leukemia virus gag gene products. In the present report, the molecular mechanisms responsible for their differential expression of gag gene products have been analyzed. Three WSV RNA genomes were shown to possess sedimentation coefficients consistent with the differences demonstrated in their allotments of helper viral sequences. The WSV variant (WSV clone 9) that expressed no detectable proteins was shown to contain the largest amount of helper viral information. Moreover, there was no additive hybridization of the WLV complementary DNA probe by RNA of this WSV clone and that of a WSV clone coding for several gag gene products. These results suggest that the lack of expression of gag gene products by WSV clone 9 is not due to a major deletion of helper viral gag gene sequences. Similar levels of WLV-specific RNA were demonstrated in cells nonproductively transformed by each WSV clone, arguing that the ability to express gag gene proteins was not related to the magnitude of viral RNA transcription. Taken together, the results are most consistent with a mechanism by which small deletions or point mutations in the genomes of some WSV variants result in premature termination of translation or synthesis of immunologically nonreactive gag gene proteins. The present findings have implications concerning the effects of evolutionary selective pressures on helper viral genetic information in mammalian transforming viruses.

The nucleic acid-binding proteins of bovine leukemia virus (BLV) and feline leukemia virus (FeLV) were isolated in a high state of purity with chloroform-methanol extraction followed by reversed-phase liquid chromatography. Selective solubilization and purity of BLV p12 and FeLV p10 was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The compositions and molecular weights were determined by amino acid analysis. An abundance of lysine and arginine residues along with their size identifies both BLV p12 and FeLV p10 as small basic proteins similar to well-defined type C viral nucleoproteins. NH2-terminal degradation by the semiautomated Edman method provided the sequence of the first 40 amino acids for both proteins. The putative nucleic acid binding site found in several type C viral nucleoproteins was contained within this sequence, with the most homology centered around an eight-amino acid region involving seven identical residues and one substitution. Antisera were developed in rabbits, and specificity and titers were determined by electroblotting and immunoautoradiography. By this technique, an immunological cross-reaction was found between BLV p12 and FeLV p10. The shared antigenic determinant most likely exists in the highly conserved eight-amino acid region. Although this sequence is also highly conserved in the nucleic acid-binding proteins of murine leukemia viruses, the shared antigenic determinant is not found in these or any other type C viruses tested. It is suggested that substitution of arginine (BLV p12/FeLV p10) to lysine (murine leukemia virus p10) is sufficient to elicit a change in antibody specificity.

Total RNAs from four BALB/c mouse tissues, containing mostly non-dividing cells (liver and kidney) or variable proportions of dividing cells (uterus and embryo) were analysed for sequences complementary to 3H-DNA transcripts synthesized from BALB/c endogenous. N- and X-tropic retroviruses. Extensive transcription of virogene information was detected in the tissues examined, but such transcription was found to be mostly limited to the homologous regions of the two virus genomes. No additivity of hybridization values could be detected when RNAs from two different tissues were mixed, which suggests that BALB/c mouse liver, kidney, uterus and embryo transcribe a common set of nucleic acid sequences of the homologous regions of the N- and X-tropic viral genomes, in addition to other sequences of the same region that are specific for indiviual tissues.

Cells of rat Thurzo-Svee leukemia produce C-type virus. Viral particles are shown to contain high-molecular weight RNA and RNA-dependent DNA polymerase the virus possesses mammalian gs-3 antigen and rat species-specific gs-1 antigen and thus appears to be of rat origin.

Feline immunodeficiency virus (FIV) (formerly feline T-lymphotropic lentivirus or FTLV) was first isolated from a group of cats in Petaluma, California in 1986. The virus is a typical lentivirus in gross and structural morphology. It replicates preferentially but not exclusively in feline T-lymphoblastoid cells, where it causes a characteristic cytopathic effect. The major structural proteins are 10, 17 (small gag), 28 (major core), 31 (endonuclease?), 41 (transmembrane?), 52 (core precursor polyprotein), 54/62 (reverse transcriptase?), and 110/130 (major envelope) kilodaltons in size. The various proteins are antigenically distinguishable from those of other lentiviruses, although serum from EIAV-infected horses will cross-react with some FIV antigens. Kittens experimentally infected with FIV manifest a transient (several days to 2 weeks) fever and neutropenia beginning 4 to 8 weeks after inoculation. This is associated with a generalized lymphadenopathy that persists for up to 9 months. Most cats recover from this initial phase of the disease and become lifelong carriers of the virus. Complete recovery does not occur to any extent in nature or in the laboratory setting. One experimentally infected cat died from a myeloproliferative disorder several months after infection. The terminal AIDS-like phase of the illness has been seen mainly in naturally infected cats. It appears a year or more following the initial infection in an unknown proportion of infected animals. FIV has been identified in cats from all parts of the world. It is most prevalent in high density populations of free roaming cats (feral and pet), and is very uncommon in closed purebred catteries. Male cats are twice as likely to become infected as females. Older male cats adopted as feral or stray animals are at the highest risk of infection, therefore. The infection rate among freely roaming cats rises throughout life, and reaches levels ranging from less than 1% to 12% or more depending on the area. Clinically affected cats tend to be 5 years or older at the time of hospitalization. Experimental and seroepidemiologic studies suggest that FIV is transmitted mainly by bites. Intimate, non-traumatic contact (mutual grooming, shared use of food, water and litter pans) is inefficient in transmitting the infection. In utero and venereal transmission could not be demonstrated in laboratory settings. There is no statistical linkage between FIV and feline leukemia virus (FeLV) infections in nature. The FeLV infection rate in FIV-infected animals is the same as it is for non-FIV-infected cats.(ABSTRACT TRUNCATED AT 400 WORDS)

Two proteins, termed gp60 and p30, have been purified to homogeneity from bovine leukemia virus (BLV) using controlled pore glass and reverse-phase liquid chromatography (RPLC). gp60 was shown to be a glycoprotein by identification of glucosamine on the amino acid analyzer. Antiserum prepared to gp60 recognized in addition to gp60 a 52,000-Da polypeptide in some virus preparations, but did not cross-react with p30. The amino and carboxyl termini of gp60 were found to be tryptophan and arginine, respectively, and a 38-residue amino-terminal sequence of gp60 (NH2TrpArgXSerLeuSerLeuGlyAsnGlnGlnTrpMetThrAlaTyrAsnGlnGluAlaLys PheSerIleSerIleAspGlnIleLeuGluAlaHisAsnGlnSerProPhe-) was obtained. A 12-residue amino-terminal sequence for p30 (NH2SerProValAlaAlaLeuThrLeuGlySerAlaLeu) was also obtained. The p30 sequence showed substantial homology to the transmembrane proteins of both types B and C retroviruses and also to a deduced sequence of the 3' region of the env gene of human T-cell leukemia virus. From these results and from elution behavior of these proteins on RPLC, it was concluded that gp60 and p30 are the BLV env gene-encoded surface glycoprotein and transmembrane protein, respectively.

A retrovirus has been isolated on the human T-cell line HuT 78 after cocultivation of a lymph node from a pig-tailed macaque (Macaca nemestrina) that had died with malignant lymphoma in 1982 at the University of Washington primate center. This isolate, designated MnIV (WPRC-1) (M. nemestrina immunodeficiency virus, Washington Primate Research Center) shows the characteristic morphology of a lentivirus and replicates to high titers in various lymphocyte lines of human and primate origin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified MnIV revealed multiple bands of structural proteins, including a major viral gag protein of 28 kilodaltons, that did not comigrate with the viral proteins of a human immunodeficiency virus (HIV [FRE-1]) that was also isolated on HuT 78 cells. The relatedness of MnIV to other lentiviruses (HTLV-III/LAV, EIAV, and visna) was examined in radioimmunoassays, by immunoblot techniques, and by N-terminal amino acid sequence analysis of the viral p28 gag protein. The immunoassays revealed cross-reactivity only between MnIV p28 and HTLV-III/LAV p24, and sequence analysis showed that 14 of the 24 N-terminal residues of MnIV p28 and HTLV-III/LAV p24 are identical. These results indicate that MnIV belongs to the same lentivirus family as HTLV-III/LAV but is only partially related to these human acquired immune deficiency syndrome retroviruses.

Proteins from bovine leukemia virus (BLV) were investigated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis in the presence and absence of 14C amino acids. Virus grown either in fetal lamb kidney (FLK) or bat lung (BL) cells revealed seven major proteins designated p10, p12, p15(1), p15(2), p24, gp30, and gp64. By tryptic peptide analysis, N-terminal amino acid analysis and radioimmunoassay it could be shown that p10, a basic protein with hydrophobic properties, is a cleavage product of p15(2), the acidic and slightly hydrophobic phosphoprotein of BLV. The structural protein p12 was shown to be a phosphorylated protein identifying it as a second major viral protein to contain phosphorous. Investigation of [3H]glucosamine incorporation, N-terminal amino acid analysis and hydrophobic properties by chloroform-methanol extraction confirmed properties of gp30 and gp64 predicted by nucleotide sequence data. The two p15 proteins have been characterized in more detail.

Quantitation by enumeration of virion particles, measurement of absorbancy at 260 nm, and densitometry on sodium dodecyl sulfate-polyacrylamide gels has shown that mouse L-M cells yielded 7- to 10-fold less endogenous C-type virions than the parental lines, L or L929. The previously noted stimulation of L-M cell virion production by a concomitant 10% increase in fetal calf serum concentration (D. A. Kindig, R. Karp, and W. H. Kirsten, 1968), was not observed.

Two species of glycosylated type C viral core polyprotein were identified on the surface of AKR spontaneous leukemia cells. One of these cell surface polyproteins was shown by immunoprecipitation to have antigenic determinants of murine leukemia virus p30, p15, p12, and p10; the other had murine leukemia virus p30, p15, and p12, but not p10, determinants. Both species were also expressed on thymocytes from 6-month-old, preleukemic AKR mice.

Peripheral leukocytes or lymph node cells and blood plasma samples from patients with lymphoid malignancies were investigated for immunological markers of BaEV, GaLV and HTLV. Antigens and antibodies were shown with radioimmunoassay. Antigen related to the p30 core protein of BaEV could be detected in each cell type of leukaemias and lymphomas. Antigen related to the GaLV p30 was found mainly in B- and O-cell forms, while that related to the p24 protein of HTLV could be detected only in two T-cell malignancies. Antibodies reactive with these antigens showed a similar distribution.

New features in the architecture of mammalian type C viruses, in particular knoblike surface projections and hexagonally arranged subunits on the core shell could be demonstrated by electron microscopy, taking advantage of newly developed preparation techniques. As examples, murine leukemia viruses (MuLVs) and newly isolated porcine and bovine C viruses are presented. The major proteins of a MuLV were isolated and partially characterized in chemical terms and with respect to their serological and other biological activities, such as interfering and hemagglutinating (HA) capacity. Most of the characterized proteins could be localized in particular substructures of the virion either by selective removal or isolation of electron microscopically identifiable constituents. The information obtained allowed the design of a more detailed model of mammalian C viruses. Special attention was devoted to the further characterization of interspecies antigens of mammalian C viruses. Different antigenic determinants were revealed. Their distribution allows further subgrouping of mammalian C viruses.

The specificity and quantitation of the rescue of RNA sequences by mammalian type C viruses has been investigated. Type C virus can package with specificity only type C viral RNA. Type C viruses do not encapsidate with comparable efficiency either type B viral or cellular globin mRNA. Conversely, a non-type C mammalian retravirus, MP-MV, cannot encapsidate type C RNA. A revertant of Kirsten sarcoma virus (Ki-SV)-transformed nonproducer cells which fails to rescue biologically active Ki-SV after superinfection with helper virus had no detectable intracellular Ki-SV-specific RNA. The results suggest specific mechanisms by which type C viral proteins can package type C viral RNA and provide an approach to classifying RNA of potentially defective endogenous retraviruses as type C in origin.

Low-infectious, nontransforming type C virus was isolated from an in vitro spontaneously transformed ST/a mouse cell line, ST-L1. The virus released by ST-L1 cells was NB-tropic and XC(-). It gave rise to very small peroxidase antibody plaques (PAP) in cultures which initially were nonproducing. Sodium dodecyl sulfate (SDS)-polyacrylamide gels of the structural proteins of the ST-L1 virus showed an envelope glycoprotein with an apparent mass of 65 kilodaltons (kdal). The mouse cells SC-1, BALB/3T3, and NIH/3T3 could be productively infected with cell-free supernatants from the ST-L1 cell line; however, virus was detected in supernatant fluids only after two to four subcultures of the infected cells. The virus thus produced was XC(+) and a large plaque former. The virus released from infected SC-1 cells was N-tropic, whereas the viruses from infected NIH/3T3 and BALB/3T3 cells were NB-tropic. The structural proteins of the N- and NB-tropic viruses could be distinguished on SDS polyacrylamide gels, the major dissimilarity being a difference in the mobility of the p30. All these viruses had an envelope glycoprotein with an apparent mass of 70 kdal. The infectivity of the viruses, measured as PAP per nanogram of p30, was 30- to 60-fold lower for the virus released from the ST-L1 cell line than that of the viruses after passage in SC-1, NIH/3T3, and BALB/3T3 cells. None of the viruses could infect rabbit or mink cells. Inoculation of the viruses into newborn mice showed that the ST-L1 virus was non-leukemogenic, whereas the NB-tropic virus selected from this after passage in BALB/3T3 or NIH/3T3 cells was highly leukemogenic. Viruses isolated from leukemic animals were indistinguishable with respect to host range and protein mobilities in SDS gels from the ones with which the mice were inoculated. Although the SC-1-selected virus was highly infectious in vitro, it was only weakly, if at all, leukemogenic.