Quantitative Changes of Some Cellular Polypeptides in C3H Mouse Cells Following Transformation by Moloney Sarcoma Virus

1983 ◽  
pp. 445-455
Author(s):  
Jes Forchhammer
Keyword(s):  
Sarcoma Virus ◽  
Mouse Cells ◽  
Moloney Sarcoma Virus ◽  
Quantitative Changes

Cellular Moloney murine sarcoma (c-mos) sequences are hypermethylated and transcriptionally silent in normal and transformed rodent cells

1982 ◽  
Vol 2 (1) ◽  
pp. 42-51
Author(s):  
S Gattoni ◽  
P Kirschmeier ◽  
I B Weinstein ◽  
J Escobedo ◽  
D Dina
Keyword(s):  
Cell Lines ◽  
Blot Hybridization ◽  
Cell Types ◽  
Single Copy ◽  
Transformed Cell Line ◽  
Murine Sarcoma Virus ◽  
Sarcoma Virus ◽  
Blotting Technique ◽  
Moloney Sarcoma Virus ◽  
Murine Sarcoma

Moloney murine sarcoma virus carries an oncogenic sequence (v-mos) which is homologous to a single copy gene (c-mos) present in the normal cells of several vertebrate species. Because of the possible significance of c-mos sequences in normal development and malignant transformation induced by physical or chemical agents, we have examined the state of integration, methylation, and transcriptional activity of c-mos sequences in a variety of normal rodent tissues, normal cell lines, or cell lines transformed by radiation or chemical carcinogens. DNA-DNA hybridization, utilizing the Southern blotting technique and a plasmid-derived DNA probe representing the v-mos sequence, gave no evidence for rearrangements of the c-mos sequence in the DNAs obtained from these diverse cell types. Parallel studies employing the restriction enzyme isoschizomers HpaII and MspI indicated that in all of these cell types the c-mos sequences were heavily methylated. In addition, analysis of cellular RNAs by blot hybridization with the v-mos probe failed to detect evidence of transcription of the c-mos sequences in any of these cell types. This was in contrast to a Moloney sarcoma virus-transformed cell line in which we found that the integrated v-mos sequence was both undermethylated and extensively transcribed. Thus, it would appear that c-mos sequences do not play a role in the transformation of rodent cells by chemical or physical agents, although the possible role of other endogenous onc sequences remains to be determined.

Mouse cells contain two distinct ras gene mRNA species that can be translated into a p21 onc protein

1982 ◽  
Vol 2 (11) ◽  
pp. 1339-1345
Author(s):  
R W Ellis ◽  
D DeFeo ◽  
M E Furth ◽  
E M Scolnick
Keyword(s):  
Normal Mouse ◽  
Velocity Sedimentation ◽  
Ras Gene ◽  
Mrna Species ◽  
P21 Ras ◽  
Murine Sarcoma Virus ◽  
Sarcoma Virus ◽  
Mouse Cells ◽  
Murine Sarcoma

The Kirsten (Ki) and Harvey (Ha) strains of murine sarcoma virus encode a 21,000-dalton protein (p21 ras) which is the product of the transforming gene of these viruses. Normal cells express low levels of p21 ras encoded by cellular genes (Ki-ras and Ha-ras) homologous to the Ki and Ha murine sarcoma virus transformation genes. A bone marrow-derived mouse cell line, 416B, has been shown to express unusually high levels of p21 ras. In this manuscript, we investigated the molecular biology of p21 ras gene expression in 416B and other normal mouse cells. We identified four distinct polyadenylated and polysome-associated RNAs, two related to Ki-ras and two to Ha-ras. The levels in 416B cells of the two Ki-ras RNAs, sized 5.2 and 2.0 kilobases, were both elevated approximately 25-fold over levels found in normal mouse cells; there was no corresponding change in 416B cells in the levels of the two Ha-ras RNAs. We partially purified the two Ki-ras mRNAs and separated them by velocity sedimentation in sucrose density gradients. Both the 5.2- and 2.0-kilobase mRNAs could be translated in vitro into p21 ras. These results show that a cellular onc protein can be translated from two distinct cellular mRNA species.

Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells

1984 ◽  
Vol 4 (12) ◽  
pp. 2929-2931 ◽  
Author(s):  
K Blochlinger ◽  
H Diggelmann
Keyword(s):  
Selectable Marker ◽  
Dna Transfer ◽  
Direct Selection ◽  
Regulatory Sequences ◽  
Hygromycin B ◽  
Dominant Marker ◽  
Dna Coding ◽  
Sarcoma Virus ◽  
Moloney Sarcoma Virus ◽  
Selection For

The DNA coding sequence for the hygromycin B phosphotransferase gene was placed under the control of the regulatory sequences of a cloned long terminal repeat of Moloney sarcoma virus. This construction allowed direct selection for hygromycin B resistance after transfection of eucaryotic cell lines not naturally resistant to this antibiotic, thus providing another dominant marker for DNA transfer in eucaryotic cells.

The long terminal repeat of an endogenous intracisternal A-particle gene functions as a promoter when introduced into eucaryotic cells by transfection

1984 ◽  
Vol 4 (10) ◽  
pp. 2128-2135
Author(s):  
K K Lueders ◽  
J W Fewell ◽  
E L Kuff ◽  
T Koch
Keyword(s):  
Long Terminal Repeat ◽  
Promoter Activity ◽  
Terminal Repeat ◽  
Flanking Sequence ◽  
Murine Sarcoma Virus ◽  
Sarcoma Virus ◽  
Mouse Cells ◽  
Moloney Murine Sarcoma Virus ◽  
Intracisternal A Particle ◽  
Murine Sarcoma

We describe experiments designed to determine whether an endogenous intracisternal A-particle (IAP) gene randomly selected from a mouse embryo library has the potential to be transcriptionally active. Assays for IAP gene transcription were done with permanently transformed rat cells and transiently transfected monkey and mouse cells. The rat cells, which had integrated IAP gene copies, contained IAP RNA. A start site within the IAP 5' long terminal repeat (LTR) was localized by S1 mapping. The promoter activity of the IAP LTR was also measured in cells 48 h after the introduction of recombinant plasmids in which bacterial chloramphenicol acetyl transferase (CAT) encoding sequences were under the control of the LTR. The IAP LTR promoted CAT activity in mouse and monkey cells. In mouse L-cells, the levels of CAT activity were 10 to 25% of those promoted by an analogous recombinant containing the Moloney murine sarcoma virus LTR as the promoter. In contrast to the Moloney murine sarcoma virus LTR, the IAP LTR was five- to eightfold more active in monkey cells than in mouse cells. The 5' and 3' LTRs were equally active, and promoter activity was dependent on having the orientation of the LTRs with respect to the CAT gene the same as their orientation with respect to the IAP gene. A 5'-flanking sequence containing a member of the highly repetitive R-sequence family increased CAT activity in COS cells 11-fold when present along with the LTR. Our results indicate that the LTR of an endogenous mouse IAP gene can function as an efficient promoter in heterologous as well as homologous cells.

scholarly journals Chemical determination of the m1 Moloney sarcoma virus pP60gag gene order: evidence for unique peptides in the carboxy terminus of the polyprotein.

1978 ◽  
Vol 75 (10) ◽  
pp. 4694-4698 ◽  
Author(s):  
M. K. Oskarsson ◽  
J. H. Elder ◽  
J. W. Gautsch ◽  
R. A. Lerner ◽  
G. F. Vande Woude
Keyword(s):  
Gene Order ◽  
Carboxy Terminus ◽  
Sarcoma Virus ◽  
Chemical Determination ◽  
Moloney Sarcoma Virus

scholarly journals THE EFFECT OF RIFAMPICIN, AND TWO DERIVATIVES, ON CELLS INFECTEDWITH MOLONEY SARCOMA VIRUS

1971 ◽  
Author(s):  
Melvin. Calvin ◽  
Urs R. Joss ◽  
Adeline J. Hackett ◽  
RobertB. Owens
Keyword(s):  
Sarcoma Virus ◽  
Moloney Sarcoma Virus
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