Abstract
Transcriptional regulatory mechanisms found in lentiviruses employ RNA enhancer elements called trans-activation responsive (TAR) elements. These nascent RNA stem-loops are cis-acting targets of virally encoded Tat effectors. Interactions between Tat and TAR increase the processivity of transcription complexes and lead to efficient copying of viral genomes. To study essential elements of this trans activation, peptide motifs from Tats of two distantly related lentiviruses, equine infectious anemia virus (EIAV) and human immunodeficiency virus type 1 (HIV-1), were fused to the coat protein of bacteriophage R17 and tested on the long terminal repeat of EIAV, where TAR was replaced by the R17 operator, the target of the coat protein. This independent RNA-tethering mechanism mapped activation domains of Tats from HIV-1 and EIAV to 47 and 15 amino acids and RNA-binding domains to 10 and 26 amino acids, respectively. Thus, a minimal lentivirus Tat consists of 25 amino acids, of which 15 modify viral transcription and 10 bind to the target RNA stem-loop.
MeSH Terms
Amino Acid Sequence
Base Sequence
Binding Sites
Capsid/genetics
Enhancer Elements, Genetic
Gene Expression Regulation, Viral
Genes, tat
HIV-1/genetics
Infectious Anemia Virus, Equine/genetics
Lentivirus/genetics
Molecular Sequence Data
Plasmids
RNA, Viral/genetics
Repetitive Sequences, Nucleic Acid
Sequence Homology, Nucleic Acid
Transcription, Genetic
Transcriptional Activation
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Derse D
Laboratory of Viral Carcinogenesis, National Cancer Institute, Frederick, Maryland 21702-1201.
Carvalho M
Carroll R
Peterlin B M
References (27)
27 references, click to expand
-
Human immunodeficiency virus as a prototypic complex retrovirus.
J Virol. 1991 Mar;65(3):1053-6
PMID: 1995941
-
The nusA gene protein of Escherichia coli. Its identification and a demonstration that it interacts with the gene N transcription anti-termination protein of bacteriophage lambda.
J Mol Biol. 1981 Mar 25;147(1):11-23
PMID: 6455533
-
A bulge structure in HIV-1 TAR RNA is required for Tat binding and Tat-mediated trans-activation.
Genes Dev. 1990 Aug;4(8):1365-73
PMID: 2227414
-
HIV-1 Tat protein trans-activates transcription in vitro.
Cell. 1990 Nov 16;63(4):791-802
PMID: 2225077
-
The HIV-1 Tat protein: an RNA sequence-specific processivity factor?
Cell. 1990 Nov 16;63(4):655-7
PMID: 2225069
-
RNA binding site of R17 coat protein.
Biochemistry. 1987 Mar 24;26(6):1563-8
PMID: 3297131
-
NusA protein is necessary and sufficient in vitro for phage lambda N gene product to suppress a rho-independent terminator placed downstream of nutL.
Proc Natl Acad Sci U S A. 1988 Apr;85(8):2494-8
PMID: 2965813
-
Structure, sequence, and position of the stem-loop in tar determine transcriptional elongation by tat through the HIV-1 long terminal repeat.
Genes Dev. 1989 Apr;3(4):547-58
PMID: 2470647
-
Site-directed mutagenesis of two trans-regulatory genes (tat-III,trs) of HIV-1.
Science. 1988 Feb 19;239(4842):910-3
PMID: 3277284
-
Anti-termination of transcription within the long terminal repeat of HIV-1 by tat gene product.
Nature. 1987 Dec 3-9;330(6147):489-93
PMID: 2825027
-
Phage lambda and the regulation of transcription termination.
Cell. 1988 Jan 15;52(1):5-6
PMID: 2449971
-
Nucleotide sequence analysis of equine infectious anemia virus proviral DNA.
Virology. 1987 Jun;158(2):300-12
PMID: 3035786
-
trans activation of human immunodeficiency virus type 1 is sequence specific for both the single-stranded bulge and loop of the trans-acting-responsive hairpin: a quantitative analysis.
J Virol. 1989 Dec;63(12):5501-4
PMID: 2479775
-
Activity of synthetic peptides from the Tat protein of human immunodeficiency virus type 1.
Proc Natl Acad Sci U S A. 1989 Oct;86(19):7397-401
PMID: 2552444
-
HIV-1 tat protein stimulates transcription by binding to a U-rich bulge in the stem of the TAR RNA structure.
EMBO J. 1990 Dec;9(12):4145-53
PMID: 2249668
-
Mutational analysis of the equine infectious anemia virus Tat-responsive element.
J Virol. 1991 Jul;65(7):3468-74
PMID: 1645778
-
Identification of lentivirus tat functional domains through generation of equine infectious anemia virus/human immunodeficiency virus type 1 tat gene chimeras.
J Virol. 1991 Jul;65(7):3460-7
PMID: 1645777
-
Analysis of arginine-rich peptides from the HIV Tat protein reveals unusual features of RNA-protein recognition.
Genes Dev. 1991 Feb;5(2):201-10
PMID: 1899841
-
Regulation of HIV gene expression by RNA-protein interactions.
Trends Genet. 1991 Jan;7(1):9-14
PMID: 2003337
-
Specific binding of a HeLa cell nuclear protein to RNA sequences in the human immunodeficiency virus transactivating region.
Proc Natl Acad Sci U S A. 1989 Jul;86(13):4858-62
PMID: 2544877
-
Mutational analysis of HIV-1 Tat minimal domain peptides: identification of trans-dominant mutants that suppress HIV-LTR-driven gene expression.
Cell. 1989 Jul 14;58(1):215-23
PMID: 2752420
-
HIV-1 tat trans-activation requires the loop sequence within tar.
Nature. 1988 Jul 14;334(6178):165-7
PMID: 3386755
-
Equine infectious anemia virus tat: insights into the structure, function, and evolution of lentivirus trans-activator proteins.
J Virol. 1990 Apr;64(4):1616-24
PMID: 2157047
-
Trans-activation by HIV-1 Tat via a heterologous RNA binding protein.
Cell. 1990 Aug 24;62(4):769-76
PMID: 2117500
-
Fragments of the HIV-1 Tat protein specifically bind TAR RNA.
Science. 1990 Sep 14;249(4974):1281-5
PMID: 2205002
-
Identification and characterization of a HeLa nuclear protein that specifically binds to the trans-activation-response (TAR) element of human immunodeficiency virus.
Proc Natl Acad Sci U S A. 1990 May;87(9):3624-8
PMID: 2333305
-
Regulation of expression of human immunodeficiency virus.
New Biol. 1990 Jan;2(1):20-31
PMID: 2078551