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Intramolecular integration within Moloney murine leukemia virus DNA

Shoemaker, Charles and Hoffmann, Joseph and Goff, Stephen P. and Baltimore, David (1981) Intramolecular integration within Moloney murine leukemia virus DNA. Journal of Virology, 40 (1). pp. 164-172. ISSN 0022-538X.

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By screening a library of unintegrated, circular Moloney murine leukemia virus (M-MuLV) DNA cloned in lambda phage, we found that approximately 20% of the M-MuLV DNA inserts contained internal sequence deletions or inversions. Restriction enzyme mapping demonstrated tht the deleted segments frequently abutted a long terminal repeat (LTR) sequence, whereas the inverted segments were usually flanked by LTR sequences, suggesting that many of the variants arose as a consequence of M-MuLV DNA molecules integrating within their own DNA. Nucleotide sequencing also suggested that most of the variant inserts were generated by autointegration. One of the recombinant M-MuLV DNA inserts contained a large inverted repeat of a unique M-MuLV sequence abutting an LTR. This molecule was shown by nucleotide sequencing to have arisen by an M-MuLV DNA Molecule integrating within a second M-MuLV DNA molecule before cloning. The autointegrated M-MuLV DNA had generally lost two base pairs from the LTR sequence at each junction with target site DNA, whereas a four-base-pair direct repeat of target site DNA flanked the integrated viral DNA. Nucleotide sequencing of preintegration target site DNA showed that this four-base-pair direct repeat was present only once before integration and was thus reiterated by the integration event. The results obtained from the autointegrated clones were supported by nucleotide sequencing of the host-virus junction of two cloned M-MuLV integrated proviruses obtained from infected rat cells. Detailed analysis of the different unique target site sequences revealed no obvious common features.

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Baltimore, David0000-0001-8723-8190
Additional Information:Copyright © 1981 by the American Society for Microbiology. Received 10 April 1981/Accepted 11 June 1981 We thank Michael Paskind for assistance with the DNA sequencing and Eli Gilboa for frequent and fruitful discussion. We are also grateful to Cary Queen for performing the computer analysis comparing the target site sequences. Finally, we thank Cliff Tabin for testing the infectivity of ZIP and ZAP DNA upon transfection into NIH/3T3 cells and for other assistance. This work was supported by Public Health Service grants CA-26717 and CA-14051 (core grant to S. E. Luria) from the National Cancer Institute. C.S. is a postdoctoral fellow of the National Cancer Institute. D.B. is an American Cancer Society Research Professor.
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Deposited On:10 Apr 2006
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