scholarly journals Preferential clustering of viral DNA sequences at or near the site of chromosomal rearrangement in fowl adenovirus type 1 DNA-transformed cell lines.

1987 ◽  
Vol 61 (1) ◽  
pp. 151-158 ◽  
Author(s):  
M Ishibashi ◽  
T H Yosida ◽  
H Yasue
Keyword(s):  
Cell Lines ◽  
Dna Sequences ◽  
Chromosomal Rearrangement ◽  
Adenovirus Type ◽  
Viral Dna ◽  
Transformed Cell ◽  
Fowl Adenovirus ◽  
Transformed Cell Lines

Isolation and characterization of four adenovirus type 12-transformed human embryo kidney cell lines

1984 ◽  
Vol 4 (1) ◽  
pp. 110-116
Author(s):  
J L Whittaker ◽  
P J Byrd ◽  
R J Grand ◽  
P H Gallimore
Keyword(s):  
Cell Lines ◽  
Dna Sequences ◽  
Human Embryo ◽  
Adenovirus Type ◽  
Left Hand ◽  
Transformed Cell ◽  
Isolation And Characterization ◽  
Human Embryo Kidney ◽  
Transformed Cell Lines

Four transformed cell lines were established from cultures of human embryo kidney (HEK) cells microinjected or transfected with cloned adenovirus 12 (Ad12) EcoRI-C DNA (0 through 16.5 map units of the left-hand end of the viral genome). Each cell line showed a different growth pattern. Southern blotting demonstrated that all of the cell lines contained Ad12-specific DNA sequences, but in the microinjected isolates these were at a much lower copy number than in the transfected isolate. Two cell lines (Ad12 HEK 1 and 3) appeared to contain tandemly repeated Ad12 EcoRI-C DNA fragments. Immunoprecipitation and Western blotting confirmed that Ad12 early region 1 (E1) proteins were being expressed by all four of the transformed cell lines, but indicated that E1A polypeptide expression was considerably less than E1B polypeptide expression. All of the Ad12-transformed HEK cell lines were tumorigenic when inoculated intracranially into athymic nude mice.

scholarly journals Isolation and characterization of four adenovirus type 12-transformed human embryo kidney cell lines.

1984 ◽  
Vol 4 (1) ◽  
pp. 110-116 ◽  
Author(s):  
J L Whittaker ◽  
P J Byrd ◽  
R J Grand ◽  
P H Gallimore
Keyword(s):  
Cell Lines ◽  
Dna Sequences ◽  
Human Embryo ◽  
Adenovirus Type ◽  
Left Hand ◽  
Transformed Cell ◽  
Isolation And Characterization ◽  
Human Embryo Kidney ◽  
Transformed Cell Lines

Four transformed cell lines were established from cultures of human embryo kidney (HEK) cells microinjected or transfected with cloned adenovirus 12 (Ad12) EcoRI-C DNA (0 through 16.5 map units of the left-hand end of the viral genome). Each cell line showed a different growth pattern. Southern blotting demonstrated that all of the cell lines contained Ad12-specific DNA sequences, but in the microinjected isolates these were at a much lower copy number than in the transfected isolate. Two cell lines (Ad12 HEK 1 and 3) appeared to contain tandemly repeated Ad12 EcoRI-C DNA fragments. Immunoprecipitation and Western blotting confirmed that Ad12 early region 1 (E1) proteins were being expressed by all four of the transformed cell lines, but indicated that E1A polypeptide expression was considerably less than E1B polypeptide expression. All of the Ad12-transformed HEK cell lines were tumorigenic when inoculated intracranially into athymic nude mice.

Expression of integrated viral DNA sequences outside the transforming region in eight adenovirus-transformed cell lines

1983 ◽  
Vol 739 (1) ◽  
pp. 48-56 ◽  
Author(s):  
A.W.M. Rijnders ◽  
M.W. Van Maarschalkerweerd ◽  
L. Visser ◽  
A.M.C.B. Reemst ◽  
J.S. Sussenbach ◽  
...  
Keyword(s):  
Cell Lines ◽  
Dna Sequences ◽  
Viral Dna ◽  
Transformed Cell ◽  
Transformed Cell Lines

scholarly journals Inhibition of p53 Transactivation Function by the Human T-Cell Lymphotropic Virus Type 1 Tax Protein

1998 ◽  
Vol 72 (2) ◽  
pp. 1165-1170 ◽  
Author(s):  
Cynthia A. Pise-Masison ◽  
Kyeong-Sook Choi ◽  
Michael Radonovich ◽  
Jürgen Dittmer ◽  
Seong-Jin Kim ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Lines ◽  
Virus Type ◽  
Primary Role ◽  
Wild Type ◽  
Transformed Cell ◽  
Human T Cell ◽  
Wild Type P53 ◽  
Transformed Cell Lines

ABSTRACT Human T-cell lymphotropic virus type 1 (HTLV-1) is the etiologic agent for adult T-cell leukemia. HTLV-1 transforms lymphocytes, and there is increasing evidence that the virus-encoded protein, Tax, plays a primary role in viral transformation. We have shown that wild-type p53 in HTLV-1-transformed cells is stabilized. This study was initiated to directly analyze whether the p53 in HTLV-1-transformed cell lines was transcriptionally active and to identify the viral gene product responsible for stabilization and inactivation. Transfection experiments using a p53-responsive reporter plasmid and γ-irradiation studies demonstrate that the wild-type p53 in HTLV-1-transformed cell lines is not fully active. Further, we demonstrate that the HTLV-1-transforming protein, Tax, stabilizes and inactivates p53 function. Cotransfection of Tax with p53 results in a greater than 10-fold reduction in p53 transcription activity. Using Gal4-p53 fusion proteins, we demonstrate that Tax inhibition of p53 transactivation function is independent of sequence-specific DNA binding. Moreover, Tax inhibits p53 function by interfering with the activity of the N-terminal activation domain (amino acids 1 to 52). We conclude that Tax is involved in the inactivation of p53 function and stabilization of p53 in HTLV-1-infected cells. The functional interference of p53 function by Tax may be important for transformation and leukemogenesis.

scholarly journals The E1a gene of adenovirus type 2 reduces the metastatic potential of ras-transformed rat embryo cells.

1988 ◽  
Vol 8 (7) ◽  
pp. 2984-2988 ◽  
Author(s):  
R Pozzatti ◽  
M McCormick ◽  
M A Thompson ◽  
G Khoury
Keyword(s):  
Cell Lines ◽  
Adenovirus Type ◽  
Metastatic Potential ◽  
Ras Oncogene ◽  
Rat Embryo ◽  
Transformed Cell ◽  
Embryo Cells ◽  
E1a Gene ◽  
Transformed Cell Lines

We have previously demonstrated that second-passage rat embryo cells transformed by the ras oncogene alone are both tumorigenic and highly metastatic when injected into nude mice. In contrast, rat embryo cells cotransformed with the ras oncogene and the adenovirus type 2 (Ad2) E1a gene are tumorigenic but either fail to metastasize or exhibit a very low metastatic potential. In this report, we demonstrate that transfection of the Ad2 E1a gene into four independent ras-transformed rat embryo cell lines results in a dramatic reduction in metastatic potential relative to that of the parental cell line. Transfection of cDNAs for the 12S and 13S E1a transcripts showed that the 12S cDNA was highly effective in reducing the metastatic potential of ras-transformed cell lines, while the 13S cDNA showed an effect in only one of the two cell lines tested. This effect is specific to the Ad2 E1a gene, since ras-transformed cell lines expressing the Ad12 E1a gene maintained their high metastatic potential. We hypothesize that the Ad2 E1a gene may regulate the expression of one or more cellular genes that contribute to the metastatic phenotype.

Alteration in expression of smooth muscle alpha-actin associated with transformation of rat 3Y1 cells

1990 ◽  
Vol 96 (4) ◽  
pp. 631-637
Author(s):  
M. Okamoto-Inoue ◽  
S. Taniguchi ◽  
H. Sadano ◽  
T. Kawano ◽  
G. Kimura ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Cell Lines ◽  
Adenovirus Type ◽  
Embryonic Cell ◽  
Stress Fibers ◽  
Two Dimensional Gel Electrophoresis ◽  
Transformed Cell ◽  
Smooth Muscle Alpha Actin ◽  
Transformed Cell Lines ◽  
Alpha Actin

Expression of actin was examined in a cultured rat embryonic cell line 3Y1 and transformed cell lines that originated from 3Y1. An alpha-actin in addition to cytoplasmic beta- and gamma-actins was detected in 3Y1 by two-dimensional gel electrophoresis. This alpha-actin was hardly detected at all in the transformants induced by Rous sarcoma virus, v-H-ras oncogene or adenovirus type 12, while the alpha-actin was retained in the transformed cell lines induced by N-methyl-N'-nitro-N-nitrosoguanidine or in SV40, which are cell lines of relatively low malignancy. Western and Northern blot analyses established that this alpha-actin was a smooth muscle alpha-isoform. An immunofluorescence study revealed that smooth muscle alpha-actin in 3Y1 cells is present in stress fibers. Thus, smooth muscle alpha-actin is also a component of actin stress fibers, as beta- and gamma-actins are in 3Y1 cells. An alteration in the expression of this actin isoform may be related to phenotypical changes accompanying transformation.

The E1a gene of adenovirus type 2 reduces the metastatic potential of ras-transformed rat embryo cells

1988 ◽  
Vol 8 (7) ◽  
pp. 2984-2988
Author(s):  
R Pozzatti ◽  
M McCormick ◽  
M A Thompson ◽  
G Khoury
Keyword(s):  
Cell Lines ◽  
Adenovirus Type ◽  
Metastatic Potential ◽  
Ras Oncogene ◽  
Rat Embryo ◽  
Transformed Cell ◽  
Embryo Cells ◽  
E1a Gene ◽  
Transformed Cell Lines

We have previously demonstrated that second-passage rat embryo cells transformed by the ras oncogene alone are both tumorigenic and highly metastatic when injected into nude mice. In contrast, rat embryo cells cotransformed with the ras oncogene and the adenovirus type 2 (Ad2) E1a gene are tumorigenic but either fail to metastasize or exhibit a very low metastatic potential. In this report, we demonstrate that transfection of the Ad2 E1a gene into four independent ras-transformed rat embryo cell lines results in a dramatic reduction in metastatic potential relative to that of the parental cell line. Transfection of cDNAs for the 12S and 13S E1a transcripts showed that the 12S cDNA was highly effective in reducing the metastatic potential of ras-transformed cell lines, while the 13S cDNA showed an effect in only one of the two cell lines tested. This effect is specific to the Ad2 E1a gene, since ras-transformed cell lines expressing the Ad12 E1a gene maintained their high metastatic potential. We hypothesize that the Ad2 E1a gene may regulate the expression of one or more cellular genes that contribute to the metastatic phenotype.

Notizen: Estimation by Reassociation Assay of Viral DNA Copies in Three Polyom a Virus Transformed Cell Lines

1976 ◽  
Vol 31 (3-4) ◽  
pp. 212-213
Author(s):  
Claudia Reinke ◽  
Gerhard Brandner
Keyword(s):  
Cell Lines ◽  
Viral Dna ◽  
Transformed Cell ◽  
Dna Reassociation ◽  
Transformed Cell Lines

Abstract The number of polyomavirus genome equivalents in three transformed hamster cell lines were shown by DNA reassociation assay to be 1.3, 1.9 and 2.5 per cell.

scholarly journals Integration and expression of several molecular forms of Rous sarcoma virus DNA used for transfection of mouse cells.

1984 ◽  
Vol 4 (7) ◽  
pp. 1260-1269 ◽  
Author(s):  
P A Luciw ◽  
H Oppermann ◽  
J M Bishop ◽  
H E Varmus
Keyword(s):  
Cell Lines ◽  
Rous Sarcoma Virus ◽  
Linear Molecule ◽  
Viral Dna ◽  
Transformed Cell ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Sarcoma Virus ◽  
Mouse Cells ◽  
Transformed Cell Lines

To assess the factors required for integration and expression of retroviral DNA, we have examined viral DNA, RNA, and protein in NIH/3T3 mouse cells transformed by transfection with various forms of cloned Rous sarcoma virus (RSV) DNA. Linear RSV DNA molecules, derived from circular DNA containing two long terminal repeats (LTRs) and permuted by cleavage at the SacI restriction endonuclease site in the leader sequence, were integrated near the ends of the linear molecule, with the LTRs on the 3' side of the src gene. Integration of a subgenomic RSV DNA fragment containing the viral src gene without intact LTRs also occurred near the ends of the linear molecule. Head-to-tail tandem arrays of RSV DNA species were observed in some transformed cell lines that received fully digested DNA and in all cell lines that received DNA ligated to produce oligomers before transfection. Closed circular RSV DNA, with one or two LTRs, integrated without apparent specificity within several regions of the viral genome. After transfection with SacI-permuted RSV DNA still linked to arms of the lambda bacteriophage vector DNA, bacteriophage sequences were joined to host DNA. Transformed cell lines produced by transfection with the various forms of RSV DNA produced similar levels of viral src protein, although the efficiency of successful transformation varied by at least two orders of magnitude. Analyses of viral polyadenylated RNA, together with the patterns of viral DNA in transformed cells, indicated that viral DNA can be integrated and expressed without regard to LTR sequences, with adjacent host DNA presumably supplying signals required for the promotion and processing of functional src mRNA.

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