Document 1016
 DOCN  M9541016
 TI    Analysis of HIV type 1 reverse transcriptase expression in a human cell
       line.
 DT    9504
 AU    Ansari-Lari MA; Gibbs RA; Department of Molecular and Human Genetics,
       Baylor College of; Medicine, Houston, Texas 77030.
 SO    AIDS Res Hum Retroviruses. 1994 Sep;10(9):1117-24. Unique Identifier :
       AIDSLINE MED/95127294
 AB    The functional analysis of human immunodeficiency virus type-1 (HIV-1)
       reverse transcriptase (RT) subunits on transient and constitutive
       expression, in the absence or presence of the HIV-1 protease (PR)
       expression, in a human cell line is described. HIV-1 RT is a heterodimer
       composed of a 51-kDa subunit (p51) and a 66-kDa subunit (p66). Cloning
       and expression of the RT region of the HIV-1 pol gene in the HT-1080
       human fibrosarcoma cell line yielded p66 without any detectable p51 and
       a low level of RT activity could be measured. Transient expression of PR
       and RT in cis generated p51 and p66, but when RT and PR were expressed
       in trans only p66 was produced. Attempts to establish a stable cell line
       expressing the PR-RT region of the pol gene were hampered by an apparent
       intolerance of HT-1080 cells to the HIV-1 PR expression. Therefore, to
       generate p51 independent of PR expression, the 51-kDa subunit was cloned
       separately. p51 lacked detectable RT activity. Coexpression of p51 and
       p66 resulted in a dramatic increase in RT activity. Stable HT-1080 cells
       producing both p51 and p66 exhibited on average a 15-fold increase in RT
       activity compared to the parental cell line. Immunofluorescence revealed
       a diffuse cytoplasmic localization of p51 and p66. To date, this is the
       first example of a human cell line that is constitutively expressing
       HIV-1 RT in the absence of HIV-1 infection.
 DE    Base Sequence  Cell Line  Cloning, Molecular  DNA Primers  Fibrosarcoma
       Fluorescent Antibody Technique  Genes, pol  Human  HIV
       Protease/BIOSYNTHESIS  HIV-1/*ENZYMOLOGY/GENETICS  Macromolecular
       Systems  Molecular Sequence Data  Molecular Weight  Polymerase Chain
       Reaction  Recombinant Proteins/BIOSYNTHESIS  Restriction Mapping
       Reverse Transcriptase/*BIOSYNTHESIS  Support, U.S. Gov't, P.H.S.
       Transfection  Tumor Cells, Cultured  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).