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Science 26 August 1988:
Vol. 241. no. 4869, pp. 1064 - 1069
DOI: 10.1126/science.2457948
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Articles

Science, Vol 241, Issue 4869, 1064-1069
Copyright © 1988 by American Association for the Advancement of Science

articles

Ordered interstrand and intrastrand DNA transfer during reverse transcription

AT Panganiban and D Fiore

McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.

Retroviruses contain two copies of the plus stranded viral RNA genome. As a means of determining whether both of these RNA's are used in the reverse transcription reaction, cells were infected with heterozygous virus particles that varied in nucleotide sequence at two separate locations at the RNA termini. The DNA proviruses formed from a single cycle of reverse transcription were then examined. Of the 12 proviruses that were characterized, all exhibited long terminal repeats (LTR's) that would be expected to arise only if both RNA templates were used for the generation of minus strand DNA. In contrast, only a single minus strand DNA appeared to be used as template for the plus strand DNA in the generation of fully double-stranded viral DNA. These results indicate that the first strand transfer step in reverse transcription is an intermolecular event while that of the second transfer is intramolecular. Thus, retroviruses contain two functionally active RNA's, and both may be required for the generation of a single linear DNA molecule. Formation of heterozygotes during retrovirus infection would be expected to result in the efficient generation of LTR recombinants.


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Science. ISSN 0036-8075 (print), 1095-9203 (online)