Human T-cell leukemia virus type II Rex binding and activity require an intact splice donor site and a specific RNA secondary structure.
The human T-cell leukemia virus type II (HTLV-II) regulatory protein Rex augments cytoplasmic levels of unspliced gag-pol mRNA by acting through a Rex-responsive element (RxRE) in the long terminal repeat. Purified Rex protein binds to long terminal repeat mRNA. Here, using an immunobinding assay to measure the binding of Rex protein to mutated RxRE RNAs, we show that efficient Rex binding requires a stem-bulge-loop RNA secondary structure (nucleotides [nt] 465 to 500) and specific sequences both within the stem-bulge (nt 470 to 476) and within a conserved upstream splice donor site (nt 449 to 455). Rex function in a transient transfection expression system correlates with Rex protein-RxRE RNA binding. The ability of HTLV-II Rex to interact directly with the HTLV-II splice donor site suggests that HTLV-II Rex may increase expression of unspliced gag-pol mRNA, in part, by inhibiting splicing.
Black
A C
AC
Department of Medicine, UCLA School of Medicine 90024-1678.
Ruland
C T
CT
Yip
M T
MT
Luo
J
J
Tran
B
B
Kalsi
A
A
Quan
E
E
Aboud
M
M
Chen
I S
IS
Rosenblatt
J D
JD
D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D016370:Gene Products, rex / Q000235:genetics* / Q000378:metabolism; D016355:Genes, pX; D015367:Human T-lymphotropic virus 2 / Q000235:genetics*; D006801:Humans; D008961:Models, Structural; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D012326:RNA Splicing; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D014162:Transfection
D016370:Gene Products, rex; D009838:Oligodeoxyribonucleotides; D012367:RNA, Viral