In Vitro Transcription of Simian Virus 40 Sequences in SV3T3 Chromatin

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1973 Aug 1

PMID 4365372

Citations 8

Authors

S M Astrin

Affiliations

Soon will be listed here.

Abstract

The template properties of chromatin were studied in a system in which transcription of unique genes could be examined. Chromation and DNA were isolated from SV3T3 cells, a line of simian virus 40 (SV40)-transformed mouse cells, and used as templates for Escherichia coli RNA polymerase. The SV40 sequences in the RNA transcribed in vitro and in RNA isolated from the transformed cells were analyzed by hybridization to radioactive RNA transcribed from SV40 DNA. The results indicate that the same sequences of the SV40 "-" strand (DNA strand which serves as a template for early RNA) are transcribed in vitro from chromatin as are found in transformed cells. In contrast, when the DNA isolated from SV3T3 chromatin by removal of proteins is used as a template, the entire SV40 "-" strand is transcribed. It is concluded that the specific template restriction observed with chromatin is due to the presence of proteins associated with the DNA.

Citing Articles

Transcription of reconstituted simian virus 40 nucleoprotein complexes.

Hidaka H, Misumi H, Oda T J Virol. 1980; 34(3):675-83.

PMID: 6247506 PMC: 288756. DOI: 10.1128/JVI.34.3.675-683.1980.

In vitro transcription of the viral-specific sequences present in the chromatin of cells transformed by simian virus 40.

Shih T, Khoury G, Martin M Proc Natl Acad Sci U S A. 1973; 70(12):3506-10.

PMID: 4357875 PMC: 427269. DOI: 10.1073/pnas.70.12.3506.

Chromatin directed transcription of 5S and tRNA genes.

Marzluff Jr W, Huang R Proc Natl Acad Sci U S A. 1975; 72(3):1082-6.

PMID: 1093165 PMC: 432470. DOI: 10.1073/pnas.72.3.1082.

Characterization of double-stranded ribonucleic acid sequences present in the initial transcription products of rat liver chromatin.

Pays E Biochem J. 1977; 165(2):237-45.

PMID: 921747 PMC: 1164894. DOI: 10.1042/bj1650237.

Transcription in vitro of immunoglobulin kappa light chain genes in isolated mouse myeloma nuclei and chromatin.

Smith M, Huang R Proc Natl Acad Sci U S A. 1976; 73(3):775-9.

PMID: 815908 PMC: 336001. DOI: 10.1073/pnas.73.3.775.

References

Gelb L, Kohne D, Martin M . Quantitation of Simian virus 40 sequences in African green monkey, mouse and virus-transformed cell genomes. J Mol Biol. 1971; 57(1):129-45. DOI: 10.1016/0022-2836(71)90123-9. View

Westphal H . SV40 DNA strand selection by Escherichia coli RNA polymerase. J Mol Biol. 1970; 50(2):407-20. DOI: 10.1016/0022-2836(70)90201-9. View

Smith K, Church R, McCarthy B . Template specificity of isolated chromatin. Biochemistry. 1969; 8(11):4271-7. DOI: 10.1021/bi00839a007. View

Bonner J, Dahmus M, Fambrough D, Huang R, Marushige K, Tuan D . The Biology of Isolated Chromatin: Chromosomes, biologically active in the test tube, provide a powerful tool for the study of gene action. Science. 1968; 159(3810):47-56. DOI: 10.1126/science.159.3810.47. View

Dawid I, WOLSTENHOLME D . Ultracentrifuge and electron microscope studies on the structure of mitochondrial DNA. J Mol Biol. 1967; 28(2):233-45. DOI: 10.1016/s0022-2836(67)80006-8. View

Seligy V, Miyagi M . Studies of template activity of chromatin isolated from metabolically active and inactive cells. Exp Cell Res. 1969; 58(1):27-34. DOI: 10.1016/0014-4827(69)90113-x. View

Zimmerman S, Sandeen D . The ribonuclease activity of crystallized pancreatic deoxyribonuclease. Anal Biochem. 1966; 14(2):269-77. DOI: 10.1016/0003-2697(66)90137-0. View

Burgess R . A new method for the large scale purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase. J Biol Chem. 1969; 244(22):6160-7. View

Paul J, Gilmour R . Restriction of deoxyribonucleic acid template activity in chromatin is organ-specific. Nature. 1966; 210(5040):992-3. DOI: 10.1038/210992a0. View

10.

Watkins J, DULBECCO R . Production of SV40 virus in heterokaryons of transformed and susceptible cells. Proc Natl Acad Sci U S A. 1967; 58(4):1396-403. PMC: 223937. DOI: 10.1073/pnas.58.4.1396. View

11.

Sambrook J, Westphal H, Srinivasan P, DULBECCO R . The integrated state of viral DNA in SV40-transformed cells. Proc Natl Acad Sci U S A. 1968; 60(4):1288-95. PMC: 224916. DOI: 10.1073/pnas.60.4.1288. View

12.

Paul J, Gilmour R . Organ-specific restriction of transcription in mammalian chromatin. J Mol Biol. 1968; 34(2):305-16. DOI: 10.1016/0022-2836(68)90255-6. View

13.

Spelsberg T, Hnilica L, ANSEVIN A . Proteins of chromatin in template restriction. 3. The macromolecules in specific restriction of the chromatin DNA. Biochim Biophys Acta. 1971; 228(2):550-62. DOI: 10.1016/0005-2787(71)90061-x. View

14.

Weinberg R, BEN-ISHAI Z, Newbold J . Poly A associated with SV40 messenger RNA. Nat New Biol. 1972; 238(82):111-3. DOI: 10.1038/newbio238111a0. View

15.

Huang R, Huang P . Effect of protein-bound RNA associated with chick embryo chromatin on template specificity of the chromatin. J Mol Biol. 1969; 39(2):365-78. DOI: 10.1016/0022-2836(69)90323-4. View

16.

Todaro G, Green H . High frequency of SV40 transformation of mouse cell line 3T3. Virology. 1966; 28(4):756-9. DOI: 10.1016/0042-6822(66)90261-3. View

17.

Sambrook J, Sharp P, Keller W . Transcription of Simian virus 40. I. Separation of the strands of SV40 DNA and hybridization of the separated strands to RNA extracted from lytically infected and transformed cells. J Mol Biol. 1972; 70(1):57-71. DOI: 10.1016/0022-2836(72)90163-5. View

18.

Tan C, Miyagi M . Specificity of transcription of chromatin in vitro. J Mol Biol. 1970; 50(3):641-53. DOI: 10.1016/0022-2836(70)90090-2. View

19.

STRAUSS Jr J, Kelly R, SINSHEIMER R . Denaturation of RNA with dimethyl sulfoxide. Biopolymers. 1968; 6(6):793-807. DOI: 10.1002/bip.1968.360060604. View

20.

Lindstrom D, DULBECCO R . Strand orientation of simian virus 40 transcription in productively infected cells. Proc Natl Acad Sci U S A. 1972; 69(6):1517-20. PMC: 426739. DOI: 10.1073/pnas.69.6.1517. View