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Analysis of a 5’ Leader Sequence on Murine Leukemia Virus 21S RNA: Heteroduplex Mapping with Long Reverse Transcriptase Products

Rothenberg, Ellen and Donoghue, Daniel J. and Baltimore, David (1978) Analysis of a 5’ Leader Sequence on Murine Leukemia Virus 21S RNA: Heteroduplex Mapping with Long Reverse Transcriptase Products. Cell, 13 (3). pp. 435-451. ISSN 0092-8674. doi:10.1016/0092-8674(78)90318-5.

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The majority of the mRNA that specifies retrovirus glycoproteins is known to be derived from the 3′ half of the genome. To examine whether the glycoprotein mRNA of murine leukemia viruses (MuLVs) might consist of portions derived from both the 5′ and 3′ ends of the viral genome, we performed hybridization with a 5′-specific probe and heteroduplex analysis with long reverse transcribed DNA. A 5′ probe was made by purifying a discrete 50 nucleotide-long reverse transcript attached to its tRNA primer. This probe was found to hybridize to RNA of the size of glycoprotein mRNA—21S, poly(A)-containing RNA—indicating that the mRNA could have a 5′ leader sequence. The 5′-specific sequences were studied by electron microscopic examination of hybrids between 21S RNA and the two longest discrete cDNA species synthesized in the endogenous reverse transcriptase reaction. One of these species, 8.8 kb long, is only made in the absence of actinomycin D, but it does not contain any self-complementary sequences, and therefore appears to be a complete transcript of the viral genome. The shorter of the two species, 8.2 kb long, is synthesized whether or not actinomycin D is present; it must terminate 500–600 nucleotides internal to the 5′ end of the template RNA. The structures observed in heteroduplexes of 21S RNA and these DNAs indicated the presence of a leader sequence approximately 500 nucleotides long at the 5′ end of the 21S RNA. Sequences comprising this leader segment in the 21S RNA mapped at the 5′ end of the genome RNA; the rest of the 21S RNA consisted of sequences from the 3′ portion of the genome. Analysis of heteroduplexes with 8.2 kb DNA suggested that actinomycin D could block the reverse transcription of most of the sequence in the genome RNA that appears as a leader in the 21S RNA.

Item Type:Article
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Rothenberg, Ellen0000-0002-3901-347X
Baltimore, David0000-0001-8723-8190
Additional Information:© 1978 by MIT. Received 8 December 1977; revised 3 January 1978; available online 28 April 2004. We are grateful to the MIT Cell Culture Facility for providing the virus which made these studies possible, and to Dr. W. A. Haseltine for communicating his results prior to publication. This research was supported by a grant from the National Cancer Institute and a contract from the Virus Cancer Program of the National Cancer Institute. D. J. D. gratefully acknowledges a fellowship from the Health Sciences Fund. D. B. is a research professor of the American Cancer Society. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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National Cancer InstituteUNSPECIFIED
Health Sciences FundUNSPECIFIED
American Cancer SocietyUNSPECIFIED
Issue or Number:3
Record Number:CaltechAUTHORS:20170207-111017078
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Official Citation:Ellen Rothenberg, Daniel J. Donoghue, David Baltimore, Analysis of a 5′ leader sequence on murine leukemia virus 21S RNA: Heteroduplex mapping with long reverse transcriptase products, Cell, Volume 13, Issue 3, March 1978, Pages 435-451, ISSN 0092-8674,
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:74134
Deposited By: Melissa Ray
Deposited On:07 Feb 2017 19:48
Last Modified:11 Nov 2021 05:25

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