Changes in synthesis of DNA binding proteins during the onset of transformation in NRK cells transformed by a temperature sensitive mutant of Rous sarcoma virus

B. E. Magun, P. H. Dorsett

Research output: Contribution to journalArticle

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Abstract

Synthesis of cytoplasmic DNA-binding proteins was investigated after a shift from the nonpermissive to the permissive temperature in NRK cells (a rat cell line) transformed by a temperature-sensitive mutant of Rous sarcoma virus [ts339(RSV)]. Cells were labeled for several generations in [3H]leucine and were pulse-labeled with [35S]methionine for 1 h at the nonpermissive temperature (39°C) and at the permissive temperature (33°C, 5 h after shift from 39°C). Proteins binding to sequential columns of double-stranded and single-stranded DNA-cellulose were examined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, and the 35S/3H ratios were obtained for each column fraction and for individual polypeptides. The protein fractions binding to single-stranded, but not double-stranded, DNA and eluting at high salt concentrations (greater than 0.60 M NaCl) showed elevated 35S/3H ratios. This indicated increased synthesis of these proteins within 5 h after the onset of transformation. The majority of the polypeptides in these fractions showed increased synthesis as a consequence of transformation. One prominent polypeptide among them constituted 0.1% of the cytosol protein and had a molecular weight of 93,000. It is concluded that the synthesis of proteins binding tightly to single-stranded DNA is increased early after the onset of transformation.

Original languageEnglish (US)
Pages (from-to)469-479
Number of pages11
JournalJournal of Virology
Volume22
Issue number2
StatePublished - 1977
Externally publishedYes

Fingerprint

Rous sarcoma virus
DNA-binding proteins
DNA-Binding Proteins
Protein Binding
polypeptides
mutants
single-stranded DNA
Temperature
synthesis
Single-Stranded DNA
protein binding
Peptides
temperature
cells
Transformed Cell Line
salt concentration
cytosol
Leucine
Sodium Dodecyl Sulfate
Methionine

ASJC Scopus subject areas

  • Immunology

Cite this

Changes in synthesis of DNA binding proteins during the onset of transformation in NRK cells transformed by a temperature sensitive mutant of Rous sarcoma virus. / Magun, B. E.; Dorsett, P. H.

In: Journal of Virology, Vol. 22, No. 2, 1977, p. 469-479.

Research output: Contribution to journalArticle

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abstract = "Synthesis of cytoplasmic DNA-binding proteins was investigated after a shift from the nonpermissive to the permissive temperature in NRK cells (a rat cell line) transformed by a temperature-sensitive mutant of Rous sarcoma virus [ts339(RSV)]. Cells were labeled for several generations in [3H]leucine and were pulse-labeled with [35S]methionine for 1 h at the nonpermissive temperature (39°C) and at the permissive temperature (33°C, 5 h after shift from 39°C). Proteins binding to sequential columns of double-stranded and single-stranded DNA-cellulose were examined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, and the 35S/3H ratios were obtained for each column fraction and for individual polypeptides. The protein fractions binding to single-stranded, but not double-stranded, DNA and eluting at high salt concentrations (greater than 0.60 M NaCl) showed elevated 35S/3H ratios. This indicated increased synthesis of these proteins within 5 h after the onset of transformation. The majority of the polypeptides in these fractions showed increased synthesis as a consequence of transformation. One prominent polypeptide among them constituted 0.1{\%} of the cytosol protein and had a molecular weight of 93,000. It is concluded that the synthesis of proteins binding tightly to single-stranded DNA is increased early after the onset of transformation.",
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N2 - Synthesis of cytoplasmic DNA-binding proteins was investigated after a shift from the nonpermissive to the permissive temperature in NRK cells (a rat cell line) transformed by a temperature-sensitive mutant of Rous sarcoma virus [ts339(RSV)]. Cells were labeled for several generations in [3H]leucine and were pulse-labeled with [35S]methionine for 1 h at the nonpermissive temperature (39°C) and at the permissive temperature (33°C, 5 h after shift from 39°C). Proteins binding to sequential columns of double-stranded and single-stranded DNA-cellulose were examined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, and the 35S/3H ratios were obtained for each column fraction and for individual polypeptides. The protein fractions binding to single-stranded, but not double-stranded, DNA and eluting at high salt concentrations (greater than 0.60 M NaCl) showed elevated 35S/3H ratios. This indicated increased synthesis of these proteins within 5 h after the onset of transformation. The majority of the polypeptides in these fractions showed increased synthesis as a consequence of transformation. One prominent polypeptide among them constituted 0.1% of the cytosol protein and had a molecular weight of 93,000. It is concluded that the synthesis of proteins binding tightly to single-stranded DNA is increased early after the onset of transformation.

AB - Synthesis of cytoplasmic DNA-binding proteins was investigated after a shift from the nonpermissive to the permissive temperature in NRK cells (a rat cell line) transformed by a temperature-sensitive mutant of Rous sarcoma virus [ts339(RSV)]. Cells were labeled for several generations in [3H]leucine and were pulse-labeled with [35S]methionine for 1 h at the nonpermissive temperature (39°C) and at the permissive temperature (33°C, 5 h after shift from 39°C). Proteins binding to sequential columns of double-stranded and single-stranded DNA-cellulose were examined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, and the 35S/3H ratios were obtained for each column fraction and for individual polypeptides. The protein fractions binding to single-stranded, but not double-stranded, DNA and eluting at high salt concentrations (greater than 0.60 M NaCl) showed elevated 35S/3H ratios. This indicated increased synthesis of these proteins within 5 h after the onset of transformation. The majority of the polypeptides in these fractions showed increased synthesis as a consequence of transformation. One prominent polypeptide among them constituted 0.1% of the cytosol protein and had a molecular weight of 93,000. It is concluded that the synthesis of proteins binding tightly to single-stranded DNA is increased early after the onset of transformation.

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