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D R Joseph

Publications and source records attributed to D R Joseph.

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Genetic linkage of C3H/HeJ and BALB/c endogenous ecotropic C-type viruses to phosphoglucomutase-1 on chromosome 5.

The genetic linkage of the endogenous C3H/HeJ C-type ecotropic virus to phosphoglucomutase-1 (0.28, recombinant fraction) on chromosome 5 was established by means of serological assays of backcrossed mice. With a combination of serological techniques and DNA-DNA hybridization the BALB/c endogenous ecotropic virus was shown to be either closely linked or allelic with the C3H/HeJ locus.

Alleles

Expression of AKR murine leukemia virus gp71-like and BALB(X) gp-71-like antigens in normal mouse tissues in the absence of overt virus expression.

By competition radioimmune assays with antisera against AKR murine leukemia virus (MuLV) gp 71 or antisera against xenotropic virus, and iodinated AKR MuLV gp71 or BALB(X) gp71, antigens serologically indistinguishable from the viral antigens can be detected in tissues of normal mice in the absence of overt virus expression. An antigen serologically indistinguishable from AKR MuLV gp71 can be readily detected in normal bone marrow cells of the common strains of mice including NIH Swiss, 129/J, and SWR/J, as well as in Mus cervicolor and Mus musculus casteneus. In contrast, this antigen is not detected in normal spleen, thymus, lymph nodes, or serum. Similarly, an antigen serologically indistinguishable from BALB(X) gp71 was found in all normal mouse sera examined. This antigen was not present in fetal liver, perfused adult liver, thymus, spleen, lymph nodes, or bone marrow of the mice examined. An equivalent antigen was detected in sera from Mus musculus casteneus but not in sera from Mus cervicolor.

AKR murine leukemia virus

Radiation leukemia in C57BL/6 mice. II. Lack of ecotropic virus expression in the majority of lymphomas.

The expression of endogenous ecotropic viruses in radiation-induced thymomas of C57BL/6 mice was examined. Competition radioimmunoassays for AKR MuLV gp71, p30, and p12 were used for viral antigen expression. 3 of 40 lymphomas had readily detectable ecotropic gp71 at levels of 95-689 ng/mg protein; the remainder of the tumors had no detectable gp71 (less than 1.0 ng/mg protein). 30 thymomas were characterized by the presence of MuLV p30 at levels of 1-10 ng/mg protein, levels that were comparable to those found in thymus extracts from age-matched, nonirradiated control. 10 tumors were characterized by having p30 levels of 10-30 ng/mg protein. In one tumor significant levels of AKR MuLV p12 were detectable. Since B-tropic and N-tropic viruses from C57BL/6 mice have glycoproteins (gp71) indistinguishable from AKR MuLV gp71 and the N-tropic virus had a p12 serologically identical to AKR MuLV p12, these results demonstrate that overt endogenous B-tropic virus was detectable in 2 of 40 thymomas and endogenous N-tropic virus was detectable in 1 of 40 thymomas. The lack of overt expression of gp71 or p12 was also confirmed by cytotoxicity assays using monospecific antisera to these viral proteins. Radiation-induced lymphomas were also examined for the presence of reverse transcriptase after chromatography of tissue extracts on poly G-Sepharose. One tumor, which was characterized by the lack of gp71, also had no detectable reverse transcriptase; whereas one tumor with gp71 was characterized by readily detectable levels of reverse transcriptase in cellular extracts. The presence of viral RNA was examined using AKR cDNA. Low levels of RNA capable of hybridizing with AKR cDNA were found in age-matched, nonirradiated mice; these hybrids had Tm's of 72 degrees C, while hybrids with AKR MuLV 70S RNA had Tm's of 80 degrees C. In 1 of 12 thymomas the concentration of hybridizable RNA and the Tm of the hybrids were identical to control values. In 9 of 12 thymomas the concentration of hybridizable sequences increased approximately three-to fivefold and the Tm of these hybrids varied from 73 to 75 degrees C. In 1 of 12 thymomas the concentration of hybridizable sequences increased over 100-fold, hybridized completely with AKR MuLV cDNA, and the hybrids had Tm's of 79 degrees C. This thymoma was also characterized by the presence of the AKR MuLV type of gp71 and p12. One tumor was characterized by a 10-to 100-fold increase in hybridizable sequences, which only partially hybridized with AKR MuLV cDNA, and hybrids had a Tm of 73 degrees C. This tumor was characterized by the presence of AKR MuLV gp71 but not AKR MuLV p12. The results taken together demonstrate that overt endogenous ecotropic virus expression is only rarely detectable in radiation-induced thymomas of C57BL/6 mice.

AKR murine leukemia virus

Separation of cellular and viral DNA polymerases by affinity chromatography on polynucleotide-Sepharose.

Polyguanylate- and poly(2'-O-methyl)uridylate-Sepharose have been prepared for affinity chromatography of DNA polymerases of viral origin (reverse transcriptase). Both cellular DNA polymerases and reverse transcriptase bind to polyguanylate-Sepharose. The cellular polymerases can be eluted from the column between 0.32 and 0.42 M NaCl while reverse transcriptase eluted between 0.56 and 0.78 M NaCl. However, only reverse transcriptase adheres to poly(2'-O-methyl)uridylate-Sepharose and can be eluted at approximately 0.35 M NaCl. The columns were used to partially purify RNA-dependent DNA polymerase from spleens of mice infected with Rauscher leukemia virus. The enzyme preparation is about 1300-fold purified and is inhibited by antiserum prepared against purified reverse transcriptase from Rauscher leukemia virus to the same extent as the virion enzyme.

Animals