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M9620567.TXT
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1996-02-26
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Document 0567
DOCN M9620567
TI The roles of nucleolar structure and function in the subcellular
location of the HIV-1 Rev protein.
DT 9602
AU Dundr M; Leno GH; Hammarskjold ML; Rekosh D; Helga-Maria C; Olson MO;
Department of Biochemistry, University of Mississippi Medical; Center,
Jackson 39216, USA.
SO J Cell Sci. 1995 Aug;108 ( Pt 8):2811-23. Unique Identifier : AIDSLINE
MED/96034319
AB The human immunodeficiency virus 1 (HIV-1) Rev transactivator protein
plays a critical role in the regulation of expression of structural
proteins by controlling the pathway of mRNA transport. The Rev protein
is located predominantly in the nucleoli of HIV-1 infected or
Rev-expressing cells. Previous studies demonstrated that the Rev protein
forms a specific complex in vitro with protein B23 which is suggested to
be a nucleolar receptor and/or carrier for the Rev protein. To study the
role of the nucleolus and nucleolar proteins in Rev function,
transfected COS-7 or transformed CMT3 cells expressing the Rev protein
were examined for subcellular locations of Rev and other proteins using
indirect immunofluorescence and immunoelectron microscopy. One day after
transfection the Rev protein was found in most cells only in the
nucleolar dense fibrillar and granular components where it colocalized
with protein B23. These were designated class 1 cells. In a second class
of cells Rev and B23 accumulated in the nucleoplasm as well as in
nucleoli. Treatment of class 1 cells with actinomycin D (AMD) under
conditions that blocked only RNA polymerase I transcription caused Rev
to completely redistribute from nucleoli to the cytoplasm.
Simultaneously, protein B23 was partially released from nucleoli, mostly
into the nucleoplasm, with detectable amounts in the cytoplasm. In cells
recovering from AMD treatment in the presence of cycloheximide Rev and
B23 showed coincident relocation to nucleoli. Class 2 cells were
resistant to AMD-induced Rev redistribution. Selective inhibition of RNA
polymerase II transcription by alpha-amanitin or by DRB did not cause
Rev to be released into the cytoplasm suggesting that active
preribosomal RNA transcription is required for the nucleolar location of
Rev. However, treatment with either of the latter two drugs at higher
doses and for longer times caused partial disruption of nucleoli
accompanied by translocation of the Rev protein to the cytoplasm. These
results suggest that the nucleolar location of Rev depends on continuous
preribosomal RNA transcription and a substantially intact nucleolar
structure.
DE Animal Antibodies Antibodies, Monoclonal Cell Line Cell
Nucleolus/*PHYSIOLOGY/*ULTRASTRUCTURE Cercopithecus aethiops
Dactinomycin/PHARMACOLOGY Gene Products, rev/ANALYSIS/DRUG
EFFECTS/*METABOLISM Human HIV-1/*METABOLISM Microscopy, Fluorescence
Microscopy, Immunoelectron Support, U.S. Gov't, P.H.S. JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).