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J Virol, August 1998, p. 6504-6510, Vol. 72, No. 8
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Human T-Cell Leukemia Virus Type 1 Reverse Transcriptase (RT) Originates from the pro and pol Open Reading Frames and Requires the Presence of RT-RNase H (RH) and RT-RH-Integrase Proteins for Its Activity

Bernadette Trentin, Nicole Rebeyrotte, and Robert Z. Mamoun*

Laboratoire Rétrovirus et Thérapie, IFR INSERM/CNRS No. 66 Pathologies Infectieuses, Université Victor Segalen Bordeaux 2, F-33076 Bordeaux Cedex, France

Received 12 December 1997/Accepted 24 April 1998

The first description of an active form of a recombinant human T-cell leukemia virus type 1 (HTLV-1) reverse transcriptase (RT) and subsequent predictions of its amino acid sequence and quaternary structure are reported here. By using amino acid alignment methods, the NH2 and COOH termini of the RT, RNase H (RH), and integrase (IN) domains of the Pol polyprotein were determined. The HTLV-1 RT seems to be unique since its NH2 terminus is probably encoded by the pro open reading frame (ORF) fused downstream, via a transframe peptide, to the polypeptide encoded by the pol ORF. The HTLV-1 Pol amino acid sequence was revealed to be highly similar to that of Rous sarcoma virus (RSV), particularly at the RT-RH hinge region. These two domains remain linked for RSV; this may also be the case for HTLV-1. In light of these results, RT, RT-RH, and RT-RH-IN genes were constructed and introduced into His-tagged protein expression vectors. The corresponding proteins were synthesized in vitro, and the DNA polymerase activities of different protein combinations were tested. Solely the RT-RH-RT-RH-IN combination was found to have a significant activity level. Velocity sedimentation analysis suggested that the HTLV-1 RT-RH and RT-RH-IN monomers are likely associated in an oligomeric structure, probably of the alpha 3/beta type.

* Corresponding author. Mailing address: B.P. 12, Université Victor Segalen Bordeaux 2, 146 Rue Léo Saignat, F-33076 Bordeaux Cedex, France. Phone: (33) 5 57 57 11 15. Fax: (33) 5 56 51 41 81. E-mail: robert.mamoun{at}retrovirether.u-bordeaux2.fr.

J Virol, August 1998, p. 6504-6510, Vol. 72, No. 8
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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