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M94A0632.TXT
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1994-10-21
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Document 0632
DOCN M94A0632
TI Involvement of the c-myb proto-oncogene in HIV-1 gene expression.
DT 9412
AU Churchill MJ; Ramsay RG; Deacon NJ; AIDS Molecular Biology Unit, NCHVR,
Macfarlane Burnet Centre,; Parkville, Australia.
SO Annu Conf Australas Soc HIV Med. 1993 Oct 28-30;5:68 (abstract no. FB3).
Unique Identifier : AIDSLINE ASHM5/94349023
AB Recently it has been shown that the HIV-1 LTR contains a high affinity
Myb-binding site, (Dasgupta et al, PNAS 1990; 87:8090). The
proto-oncogene c-myb, is expressed in many cell types including
hemopoietic cell lines and tissues where it is thought to be associated
with the regulation of proliferation and differentiation. The expression
of c-myb in different CD4+ T-cell lines was examined in both cytoplasmic
mRNA and nuclear protein. Levels of c-myb mRNA determined by Northern
blot analysis were approximately 10 fold higher in MT-2 cells (an HTLV-1
transformed line) compared with CEM cells, while unstimulated PBLs
showed a level 1/10 that of CEM cells. Examination of Myb protein by
Western blotting (WB) also showed the higher expression of c-myb in MT-2
cells. Mobility shift assays were used to investigate the DNA binding
activity of nuclear extract from uninfected and HIV-1 infected MT2 and
CEM cells. Using a synthetic Myb-binding template, we found that the
protein levels observed by WB correlate well with DNA binding activity
using both core Myb responsive element (MBS-1) and HIV-1 LTR Myb
responsive element (LTR-9) oligonucleotides. Transfection of HIV-1
infected CEM cells with c-myb expression plasmid pACTC3 increased
cotransfected HIV LTR driven reporter gene activity and HIV cytopathic
effect. Antisense oligonucleotides to c-myb mRNA inhibited infection of
MT-2 cells by HIV-1. These data suggest that the elevated levels of
c-myb expression in MT-2 cells is a requisite for the rapid kinetics of
HIV-1 infection and virus production in these cells over other T-cell
lines.
DE Cell Line, Transformed Gene Expression Regulation, Viral/PHYSIOLOGY
Human HIV-1/*GENETICS Proto-Oncogene Proteins/GENETICS Proto-Oncogene
Proteins c-myc/*GENETICS Repetitive Sequences, Nucleic Acid/*GENETICS
RNA, Messenger/GENETICS Virus Replication/GENETICS MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).