Abstract
A highly specific procedure for the isolation of deletion mutants is described. The size and location of the deletions can be predetermined. By this method a series of deletion mutants mapping within and near the untranslated 5' leader sequence of the late 16S mRNA of simian virus 40 have been isolated. The boundaries of the deletions have been accurately determined by DNA sequence analysis. The deletions range from 20 to 223 nucleotides. All these deletions mutants are viable and grow without helper virus. The largest of these deletions removes the entire leader sequence except for six nucleotides at the 3' end that are probably involved in covalent linkage with the 5' end of the body of the mRNA located 937 nucleotides away on the genome. Three of the deletion mutants remove the 5' end of the leader that normally bears the cap structure of the mRNA. A large segment immediately preceding the leader sequence is also removed in one of these mutants, ruling out the generation of the 5' end of the mRNA via initiation of transcription at this point. The circularization of linear infecting DNA producing the DNA of the deletion mutants proceeds mainly by way of blunt end ligation in vivo.
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- Aloni Y., Dhar R., Laub O., Horowitz M., Khoury G. Novel mechanism for RNA maturation: the leader sequences of simian virus 40 mRNA are not transcribed adjacent to the coding sequences. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3686–3690. doi: 10.1073/pnas.74.9.3686. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carbon J., Shenk T. E., Berg P. Biochemical procedure for production of small deletions in simian virus 40 DNA. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1392–1396. doi: 10.1073/pnas.72.4.1392. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Celma M. L., Dhar R., Pan J., Weissman S. M. Comparison of the nucleotide sequence of the messenger RNA for the major structural protein of SV40 with the DNA sequence encoding the amino acids of the protein. Nucleic Acids Res. 1977 Aug;4(8):2549–2550. [PMC free article] [PubMed] [Google Scholar]
- Cole C. N., Landers T., Goff S. P., Manteuil-Brutlag S., Berg P. Physical and genetic characterization of deletion mutants of simian virus 40 constructed in vitro. J Virol. 1977 Oct;24(1):277–294. doi: 10.1128/jvi.24.1.277-294.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Danna K. J., Sack G. H., Jr, Nathans D. Studies of simian virus 40 DNA. VII. A cleavage map of the SV40 genome. J Mol Biol. 1973 Aug 5;78(2):363–376. doi: 10.1016/0022-2836(73)90122-8. [DOI] [PubMed] [Google Scholar]
- Dhar R., Subramanian K. N., Pan J., Weissman S. M. Nucleotide sequence of a fragment of SV40 DNA that contains the origin of DNA replication and specifies the 5' ends of "early" and "late" viral RNA. IV. Localization of the SV40 DNA complementary to the 5' ends of viral mRNA. J Biol Chem. 1977 Jan 10;252(1):368–376. [PubMed] [Google Scholar]
- Dhar R., Subramanian K. N., Pan J., Weissman S. M. Structure of a large segment of the genome of simian virus 40 that does not encode known proteins. Proc Natl Acad Sci U S A. 1977 Mar;74(3):827–831. doi: 10.1073/pnas.74.3.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
- Ghosh P. K., Reddy V. B., Swinscoe J., Choudary P. V., Lebowitz P., Weissman S. M. The 5'-terminal leader sequence of late 16 S mRNA from cells infected with simian virus 40. J Biol Chem. 1978 May 25;253(10):3643–3647. [PubMed] [Google Scholar]
- Haegeman G., Fiers W. Evidence for 'splicing' of SV40 16S mRNA. Nature. 1978 May 4;273(5657):70–73. doi: 10.1038/273070a0. [DOI] [PubMed] [Google Scholar]
- Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
- Hsu M. T., Ford J. Sequence arrangement of the 5' ends of simian virus 40 16S and 19S mRNAs. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4982–4985. doi: 10.1073/pnas.74.11.4982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lai C. J., Nathans D. Deletion mutants of simian virus 40 generated by enzymatic excision of DNA segments from the viral genome. J Mol Biol. 1974 Oct 15;89(1):179–193. doi: 10.1016/0022-2836(74)90169-7. [DOI] [PubMed] [Google Scholar]
- Lavi S., Groner Y. 5'-Terminal sequences and coding region of late simian virus 40 mRNAs are derived from noncontiguous segments of the viral genome. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5323–5327. doi: 10.1073/pnas.74.12.5323. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lavi S., Shatkin A. J. Methylated simian virus 40-specific RNA from nuclei and cytoplasm of infected BSC-1 cells. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2012–2016. doi: 10.1073/pnas.72.6.2012. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maniatis T., Kee S. G., Efstratiadis A., Kafatos F. C. Amplification and characterization of a beta-globin gene synthesized in vitro. Cell. 1976 Jun;8(2):163–182. doi: 10.1016/0092-8674(76)90001-5. [DOI] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mertz J. E., Berg P. Defective simian virus 40 genomes: isolation and growth of individual clones. Virology. 1974 Nov;62(1):112–124. doi: 10.1016/0042-6822(74)90307-9. [DOI] [PubMed] [Google Scholar]
- Mertz J. E., Berg P. Viable deletion mutants of simian virus 40: selective isolation by means of a restriction endonuclease from Hemophilus parainfluenzae. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4879–4883. doi: 10.1073/pnas.71.12.4879. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Muthukrishnan S., Both G. W., Furuichi Y., Shatkin A. J. 5'-Terminal 7-methylguanosine in eukaryotic mRNA is required for translation. Nature. 1975 May 1;255(5503):33–37. doi: 10.1038/255033a0. [DOI] [PubMed] [Google Scholar]
- Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
- Roberts R. J. Restriction endonucleases. CRC Crit Rev Biochem. 1976 Nov;4(2):123–164. doi: 10.3109/10409237609105456. [DOI] [PubMed] [Google Scholar]
- Shenk T. E., Carbon J., Berg P. Construction and analysis of viable deletion mutants of simian virus 40. J Virol. 1976 May;18(2):664–671. doi: 10.1128/jvi.18.2.664-671.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Subramanian K. N., Pan J., Zain S., Weissman S. M. The mapping and ordering of fragments of SV40 DNA produced by restriction endonucleases. Nucleic Acids Res. 1974 Jun;1(6):727–752. doi: 10.1093/nar/1.6.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Subramanian K. N., Shenk T. Definition of the boundaries of the origin of DNA replication in simian virus 40. Nucleic Acids Res. 1978 Oct;5(10):3635–3642. doi: 10.1093/nar/5.10.3635. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tomassini J., Roychoudhury R., Wu R., Roberts R. J. Recognition sequence of restriction endonuclease KpnI from Klebsiella pneumoniae. Nucleic Acids Res. 1978 Nov;5(11):4055–4064. doi: 10.1093/nar/5.11.4055. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van de Voorde A., Contreras R., Rogiers R., Fiers W. The initiation region of the SV40 VP1 gene. Cell. 1976 Sep;9(1):117–120. doi: 10.1016/0092-8674(76)90057-x. [DOI] [PubMed] [Google Scholar]
- Yang R. C., Van de Voorde A., Fiers W. Cleavage map of the simian-virus-40 genome by the restriction endonuclease III of Haemopholus aegyptius. Eur J Biochem. 1976 Jan 2;61(1):101–117. doi: 10.1111/j.1432-1033.1976.tb10002.x. [DOI] [PubMed] [Google Scholar]
- Yang R., Danna K., Van de Voorde A., Fiers W. Location of the small restriction fragments, Hind-L, Hind-M and Hpa-E, on the simian virus 40 genome. Virology. 1975 Nov;68(1):260–265. doi: 10.1016/0042-6822(75)90167-1. [DOI] [PubMed] [Google Scholar]