Characterization of a Rous Sarcoma Virus Mutant Defective in Packaging Its Own Genomic RNA: Biochemical Properties of Mutant TK15 and Mutant-induced Transformants

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 1984 Jul 1

PMID 6202881

Citations 43

Authors

T Koyama

F Harada

S Kawai

Affiliations

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Abstract

The accompanying paper (S. Kawai and T. Koyama , J. Virol. 51:147-153, 1984) describes the isolation and biological properties of a mutant, TK15 , derived from a Rous sarcoma virus mutant, tsNY68 . The cis-acting defect of the mutant is analyzed biochemically in this paper. TK15 virions released from virus-producing 15c (+) cells were deficient in viral genomic 39S RNA, although comparable amounts of viral RNAs were transcribed in 15c (+) and tsNY68 -infected cells. Analysis of provirus DNA occurring in 15c (+) cells suggested that the mutant genome had a deletion of ca. 250 bases near the 5' end of the genome somewhere between the primer binding site and the 5' end of the gag-coding region. These findings indicate that at least part of the sequence lost in the TK15 genome is indispensable for packaging viral genomic RNA into virions. TK15 induces nonvirus -producing 15c (-) transformants at high frequency. Southern blot analysis of DNAs from those 15c (-) clone cells revealed that TK15 -derived proviruses contained various extents of internal deletions. Many 15c (-) clones had a provirus carrying only the src gene with long terminal repeat sequences at both ends. The mechanism for the segregation of 15c (-) cells is discussed.

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References

Wang L, Stacey D . Participation of subgenomic retroviral mRNAs in recombination. J Virol. 1982; 41(3):919-30. PMC: 256828. DOI: 10.1128/JVI.41.3.919-930.1982. View

Swanstrom R, Varmus H, Bishop J . Nucleotide sequence of the 5' noncoding region and part of the gag gene of Rous sarcoma virus. J Virol. 1982; 41(2):535-41. PMC: 256782. DOI: 10.1128/JVI.41.2.535-541.1982. View

Watanabe S, Temin H . Encapsidation sequences for spleen necrosis virus, an avian retrovirus, are between the 5' long terminal repeat and the start of the gag gene. Proc Natl Acad Sci U S A. 1982; 79(19):5986-90. PMC: 347036. DOI: 10.1073/pnas.79.19.5986. View

Pugatsch T, Stacey D . Identification of a sequence likely to be required for avian retroviral packaging. Virology. 1983; 128(2):505-11. DOI: 10.1016/0042-6822(83)90279-9. View

Kawai S, Koyama T . Characterization of a Rous sarcoma virus mutant defective in packaging its own genomic RNA: biological properties of mutant TK15 and mutant-induced transformants. J Virol. 1984; 51(1):147-53. PMC: 254412. DOI: 10.1128/JVI.51.1.147-153.1984. View

Mann R, Mulligan R, Baltimore D . Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983; 33(1):153-9. DOI: 10.1016/0092-8674(83)90344-6. View

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

Kawai S, Yamamoto T . Isolation of different kinds of non-virus producing chick cells transformed by Schmidt-Ruppin strain (subgroup A) of Rous sarcoma virus. Jpn J Exp Med. 1970; 40(4):243-56. View

Kawai S, Hanafusa H . The effects of reciprocal changes in temperature on the transformed state of cells infected with a rous sarcoma virus mutant. Virology. 1971; 46(2):470-9. DOI: 10.1016/0042-6822(71)90047-x. View

10.

Kawai S, Hanafusa H . Plaque assay for some strains of avian leukosis virus. Virology. 1972; 48(1):126-35. DOI: 10.1016/0042-6822(72)90120-1. View

11.

Aviv H, Leder P . Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972; 69(6):1408-12. PMC: 426713. DOI: 10.1073/pnas.69.6.1408. View

12.

Ikemura T, Dahlberg J . Small ribonucleic acids of Escherichia coli. I. Characterization by polyacrylamide gel electrophoresis and fingerprint analysis. J Biol Chem. 1973; 248(14):5024-32. View

13.

Sawyer R, Dahlberg J . Small RNAs of Rous sarcoma virus: characterization by two-dimensional polyacrylamide gel electrophoresis and fingerprint analysis. J Virol. 1973; 12(6):1226-37. PMC: 356763. DOI: 10.1128/JVI.12.6.1226-1237.1973. View

14.

Kawai S, Hanafusa H . Isolation of defective mutant of avian sarcoma virus. Proc Natl Acad Sci U S A. 1973; 70(12):3493-7. PMC: 427266. DOI: 10.1073/pnas.70.12.3493. View

15.

Mizutani S, Kang C, Temin H . Endogenous RNA-directed DNA polymerase activity in virions of RNA tumor viruses and in a fraction from normal chicken cells. Methods Enzymol. 1974; 29:119-24. DOI: 10.1016/0076-6879(74)29014-1. View

16.

Harada F, Sawyer R, Dahlberg J . A primer ribonucleic acid for initiation of in vitro Rous sarcarcoma virus deoxyribonucleic acid synthesis. J Biol Chem. 1975; 250(9):3487-97. View

17.

Southern E . Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975; 98(3):503-17. DOI: 10.1016/s0022-2836(75)80083-0. View

18.

Kawai S, Hanafusa H . Recombination between a temperature-sensitive mutant and a deletion mutant of Rous sarcoma virus. J Virol. 1976; 19(2):389-97. PMC: 354876. DOI: 10.1128/JVI.19.2.389-397.1976. View

19.

Strohman R, Moss P, Spector D, Przybyla A, Paterson B . Messenger RNA for myosin polypeptides: isolation from single myogenic cell cultures. Cell. 1977; 10(2):265-73. DOI: 10.1016/0092-8674(77)90220-3. View

20.

Taylor J . An analysis of the role of tRNA species as primers for the transcription into DNA of RNA tumor virus genomes. Biochim Biophys Acta. 1977; 473(1):57-71. DOI: 10.1016/0304-419x(77)90007-5. View