Abstract
The HIV-1 Tat protein regulates transcription elongation through binding to the viral TAR RNA stem-loop structure. We have isolated a novel 87 kDa cyclin C-related protein (cyclin T) that interacts specifically with the transactivation domain of Tat. Cyclin T is a partner for CDK9, an RNAPII transcription elongation factor. Remarkably, the interaction of Tat with cyclin T strongly enhances the affinity and specificity of the Tat:TAR RNA interaction, and confers a requirement for sequences in the loop of TAR that are not recognized by Tat alone. Moreover, overexpression of human cyclin T rescues Tat activity in nonpermissive rodent cells. We propose that Tat directs cyclin T-CDK9 to RNAPII through cooperative binding to TAR RNA.
MeSH Terms
3T3 Cells
Amino Acid Sequence
Animals
Bacterial Proteins/genetics
CHO Cells
Cell Nucleus/enzymology
Chemoreceptor Cells
Cloning, Molecular
Cricetinae
Cyclin C
Cyclin T
Cyclin-Dependent Kinases/metabolism
Cyclins/genetics,metabolism
DNA Footprinting
Escherichia coli Proteins
Gene Expression Regulation, Viral/physiology
Gene Products, tat/metabolism
HIV-1
Humans
Membrane Proteins/genetics
Mice
Molecular Sequence Data
Nucleic Acid Conformation
Positive Transcriptional Elongation Factor B
Protein Serine-Threonine Kinases/metabolism
RNA Polymerase II/metabolism
RNA, Viral/chemistry,metabolism
Receptors, Cell Surface/genetics
Recombinant Proteins/genetics
Ribonucleases
Substrate Specificity
Transcription, Genetic/physiology
tat Gene Products, Human Immunodeficiency Virus
Chemicals
Bacterial Proteins
CCNC protein, human
CCNT1 protein, human
Ccnc protein, mouse
Ccnt1 protein, mouse
Cyclin C
Cyclin T
Cyclins
Escherichia coli Proteins
Gene Products, tat
Membrane Proteins
RNA, Viral
Receptors, Cell Surface
Recombinant Proteins
Tar protein, E coli
tat Gene Products, Human Immunodeficiency Virus
Positive Transcriptional Elongation Factor B
Protein Serine-Threonine Kinases
Cyclin-Dependent Kinases
RNA Polymerase II
Ribonucleases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wei P
Regulatory Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037-1099, USA.
Garber M E
Fang S M
Fischer W H
Jones K A