Expression and mutational analysis of the reverse transcriptase of the lentivirus equine infectious anemia virus.
The reverse transcriptase of equine infectious anemia virus (EIAV) shows sequence similarity with the reverse transcriptases of other lentiviruses, particularly with those of human immunodeficiency viruses types 1 and 2 (HIV-1 and HIV-2). We have constructed a plasmid that when introduced into E. coli induces the synthesis of substantial quantities of the nearly authentic EIAV reverse transcriptase. The viral and bacterially expressed reverse transcriptases are similar in their molecular weights. The bacterial expression clone was used to generate deletion mutants of the protein. Mutations in both amino and carboxyl terminal regions of the polypeptide strongly affect the DNA polymerase activity of the enzyme. Thus, EIAV reverse transcriptase resembles the reverse transcriptases of HIV-1 and HIV-2 and can serve as a suitable enzyme for studying the structure-function relationship in lentiviral reverse transcriptase.
Shaharabany
M
M
Department of Cell Biology and Histology, Sackler School of Medicine, Tel Aviv University, Israel.
Rice
N R
NR
Hizi
A
A
D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D015751:Genes, pol; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology; D015498:HIV-2 / Q000201:enzymology; D004860:Infectious Anemia Virus, Equine / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D011533:Proviruses / Q000201:enzymology / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D017384:Sequence Deletion
D011994:Recombinant Proteins; C072416:reverse transcriptase, Human immunodeficiency virus 2; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase