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Journal of Virology, September 2000, p. 8775-8780, Vol. 74, No. 18
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The 17 Nucleotides Downstream from the env Gene Stop Codon Are Important for Murine Leukemia Virus Packaging

Seung Shin Yu,1,2 Jong-Mook Kim,2 and Sunyoung Kim1,2,*

ViroMed Limited, Technology Business Incubator,1 and Institute for Molecular Biology and Genetics, Seoul National University,2 Seoul 151-742, Korea

Received 20 April 2000/Accepted 10 June 2000

We have identified a previously unknown nucleotide sequence important for the packaging of murine leukemia virus. This nucleotide sequence is located downstream from the stop codon of the env gene but does not overlap the polypurine tract. Deletion of 17 bp from this region resulted in a more than 10-fold decrease in viral titer. Consistent with this result, the deletion mutant showed a 20- to 30-fold drop in the amount of virion RNA in the culture supernatant. The total amount of virion protein in the culture supernatant was comparable for the deletion mutant and the parental virus, suggesting that the mutant construct could release the empty viral particles. These results suggested that the packaging signal sequence might be present at the two extreme sites of the viral genome, one in the region around the splice donor sequence downstream from the 5' long terminal repeat (LTR) and the other immediately upstream from the 3' LTR. Implications for gene therapy, especially in regard to construction of retroviral vectors and packaging constructs, are discussed.

* Corresponding author. Mailing address: Institute for Molecular Biology and Genetics, Seoul National University, Kwan-Ak-Gu, Seoul 151-742, Korea. Phone: 82-2-880-7529. Fax: 82-2-875-0907. E-mail: sunyoung{at}plaza.snu.ac.kr.

Journal of Virology, September 2000, p. 8775-8780, Vol. 74, No. 18
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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