Comparative studies of bacterially expressed integrase proteins of caprine arthritis-encephalitis virus, maedi-visna virus and human immunodeficiency virus type 1
Integrase (IN) proteins mediate an essential step in retroviral life cycles, the integration of reverse-transcribed viral DNA into the host genome. To create tools for direct comparative investigations, hexahistidine-tagged IN proteins of the phylogenetically related lentiviruses caprine arthritis-encephalitis virus (CAEV), maedi-visna virus (MVV) and human immunodeficiency virus type 1 (HIV-1) were expressed in Escherichia coli. After purification by affinity chromatography, the active enzymes were compared in vitro for their site-specific cleavage, integration and disintegration activities on cognate and non-cognate oligonucleotide substrates. It was found that CAEV IN and MVV IN catalyse both site-specific cleavage and disintegration with high efficiencies, reduced substrate specificities and similar reaction patterns. Comparisons with the respective activities of HIV-1 IN revealed basic functional similarities as well as considerable differences such as more restricted substrate requirements for site-specific cleavage. On the other hand, all three enzymes catalyse disintegration almost independent of the substrate origin. Furthermore, MVV IN was shown to join oligonucleotides as efficiently as HIV-1 IN, albeit with reduced substrate specificity. In contrast, no detectable strand transfer activities occurred with CAEV IN.
AdachiA.,
GendblmanH. E.,
KoenigS.,
FolksT.,
WilleyR.,
RabsonA.,
MartinM. A.1986; Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone. Journal of Virology 59:284–291
BradfordM. M.1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
BraunM. J.,
ClementsJ. E.,
GondaM. A.1987; The visna virus genome: evidence for a hypervariable site in the env gene and sequence homology among lentivirus envelope proteins. Journal of Virology 61:4046–4054
BurkeC. J.,
SanyalG.,
BrunerM. W.,
RyanJ. A.,
LaFeminaR. L.,
RobbinsH. L.,
ZeftA. S.,
MiddaughC. R.,
CordingleyM. G.1992; Structural implications of spectroscopic characterization of a putative zinc finger peptide from HIV-1 integrase. Journal of Biological Chemistry 267:9639–9644
BushmanF. D.,
CraigieR.1991; Activities of human immunodeficiency virus (HIV) integration protein in vitro: specific cleavage and integration of HIV DNA. Proceedings of the National Academy of Sciences, USA 88:1339–1343
BushmanF. D.,
WangB.1994; Rous sarcoma virus integrase protein: mapping functions for catalysis and substrate binding. Journal of Virology 68:2215–2223
BushmanF. D.,
EngelmanA.,
PalmerI.,
WingfieldP.,
CraigieR.1993; Domains of the integrase protein of human immunodeficiency virus type 1 responsible for polynucleotidyl transfer and zinc binding.. Proceedings of the National Academy of Sciences, USA 90:3428–3432
CheeversW. P.,
McGuireT. C.1988; The lentiviruses: maedi/ visna, caprine arthritis-encephalitis, and equine infectious anemia. Advances in Virus Research 3:189–215
ChowS. A.,
BrownP. O.1994; Substrate features important for recognition and catalysis by human immunodeficiency virus type 1 integrase identified by using novel DNA substrates. Journal of Virology 68:3896–3907
ChowS. A.,
VincentK. A.,
EllisonV.,
BrownP. O.1992; Reversal of integration and DNA splicing mediated by integrase of human immunodeficiency virus. Science 255:723–726
CraigieR.,
FujiwaraT.,
BushmanF.1990; The IN protein of Moloney murine leukemia virus processes the viral DNA ends and accomplishes their integration in vitro. Cell 62:829–837
DrelichM.,
WilhelmR.,
MousJ.1992; Identification of amino acid residues critical for endonuclease and integration activities of HIV-1 IN protein in vitro. Virology 188:459–468
EngelmanA.,
CraigieR.1992; Identification of conserved amino acid residues critical for human immunodeficiency virus type 1 integrase function in vitro. Journal of Virology 66:6361–6369
EngelmanA.,
BushmanF. D.,
CraigieR.1993; Identification of discrete functional domains of HIV-1 integrase and their organization within an active multimeric complex. EMBO Journal 12:3269–3275
FrankR.,
& KosterH.1979; DNA chain length markers and the influence of base composition on electrophoretic mobility of oligodeoxyribonucleotides in polyacrylamide-gels.. Nucleic Acids Research 7:2069–2087
HiziA.,
HughesS. H.1988; Expression of the Moloney murine leukemia vims and human immunodeficiency virus integration proteins in Escherichia coli. Virology 167:634–638
HongT.,
MurphyE.,
GroarkeJ.,
DrlicaK.1993; Human immunodeficiency virus type 1 DNA integration: fine structure target analysis using synthetic oligonucleotides. Journal of Virology 67:1127–1131
KalpanaG. V.,
GoffS. P.1993; Genetic analysis of homomeric interactions of human immunodeficiency virus type 1 integrase using the yeast two-hybrid system. Proceedings of the National Academy of Sciences, USA 90:10593–10597
KatzR. A.,
MerkelG.,
KulkoskyJ.,
LeisJ.,
SkalkaA. M.1990; The avian retroviral IN protein is both necessary and sufficient for integrative recombination in vitro. Cell 6:87–95
KatzmanM.,
KatzR. A.,
SkalkaA. M.,
LeisJ.1989; The avian retroviral integration protein cleaves the terminal sequences of linear viral DNA at the in vivo sites of integration. Journal of Virology 63:5319–5327
KhanE.,
MackJ. P. G.,
KatzR. A.,
KulkoskyJ.,
SkalkaA. M.1991; Retroviral integrase domains: DNA binding and the recognition of LTR sequences. Nucleic Acids Research 19:851–860
KulkoskyJ.,
JonesK. S.,
KatzR. A.,
MackJ. P. G.,
SkalkaM.1992; Residues critical for retroviral integrative recombination in a region that is highly conserved among retroviral/ retrotransposon integrases and bacterial insertion sequence trans-posases. Molecular and Cellular Biology 12:2331–2338
LaFeminaR. L.,
SchneiderC. L.,
RobbinsH. L.,
CallahanP. L.,
LeGrowK.,
RothE.,
SchleifW. A.,
EminiE. A.1992; Requirement of active human immunodeficiency virus type 1 integrase enzyme for productive infection of human T-lymphoid cells. Journal of Virology 66:7414–7419
LeavittA. D.,
RoseR. B.,
VarmusH. E.1992; Both substrate and target oligonucleotide sequences affect in vitro integration mediated by human immunodeficiency virus type 1 integrase protein produced in Saccharomyces cerevisiae. Journal of Virology 66:2359–2368
McEuenA. R.,
EdwardsB.,
KoepkeK. A.,
BallA. E.,
JenningsA.,
WolstenholmeA. J.,
DansonM. J.,
HoughD. W.1992; Zinc binding by retroviral integrase. Biochemical and Biophysical Research Communications 189:813–818
MurphyJ. E.,
GoffS. P.1992; A mutation at one end of moloney murine leukemia virus DNA blocks cleavage of both ends by the viral integrase in vivo. Journal of Virology 66:5092–5095
NarayanO.,
ZinkM. C.,
GorrellM.,
CraneS.,
HusoD.,
JollyP.,
SaltarelliM.,
AdamsR. J.,
ClementsJ. E.1993; The lentiviruses of sheep and goats. In The Retroviridae vol 2 pp 229–255 Edited by
LevyJ. A.
New York:: Plenum Press.;
PahlA.,
FlugelR. M.1993; Endonucleolytic cleavages and DNA-joining activities of the integration protein of human foamy virus. Journal of Virology 67:5426–5434
SakaiH.,
KawamuraM.,
SakuragiJ.-I.,
SakuragiS.,
ShibataR.,
IshimotoA.,
OnoN.,
UedaS.,
AdachiA.1993; Integration is essential for efficient gene expression of human immunodeficiency virus type 1. Journal of Virology 67:1169–1174
SaltarelliM.,
QueratG.,
KoningsD. A. M.,
VigneR.,
ClementsJ. E.1990; Nucleotide sequence and transcriptional analysis of molecular clones of CAEV which generate infectious virus. Virology 179:347–364
SambrookJ.,
FritschE. F.,
ManiatisM.1989; Molecular Cloning: A Laboratory Manual, 2nd edn. New York: Cold Spring Harbor Laboratory. Sanger, F., Nicklen, S. & Coulson, A. R. (1977). DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, USA 74:5463–5467
SchauerM.,
BillichA.1992; The N-terminal region of HIV-1 integrase is required for integration activity, but not for DNA-binding. Biochemical and Biophysical Research Communications 185:874–880
ShermanP. A.,
DicksonM. L.,
FyfeJ. A.1992; Human immunodeficiency virus type 1 integration protein: DNA sequence requirements for cleaving and joining reactions. Journal of Virology 66:3593–3601
Van GentD. C.,
Oude GroenegerA. A. M.,
PlasterkR. H. A.1992; Mutational analysis of the integrase protein of human immunodeficiency virus type 2. Proceedings of the National Academy of Sciences, USA 89:9598–9602
Van GentD. C.,
VinkC.,
Oude GroenegerA. A. M.,
PlasterkR. H. A.1993; Complementation between HIV integrase proteins mutated in different domains. EMBO Journal 12:3261–3267
VincentK. A.,
EllisonV.,
ChowS. A.,
BrownP. O.1993; Characterization of human immunodeficiency virus type 1 integrase expressed in Escherichia coli and analysis of variants with amino-terminal mutations. Journal of Virology 67:425–437
VinkC.,
Van GentD. C.,
ElgersmaY.,
PlasterkR. H. A.1991a; Human immunodeficiency virus integrase protein requires a subterminal position of its viral DNA recognition sequence for efficient cleavage. Journal of Virology 65:4636–4644
VinkC.,
YeheskielyE.,
van der MarelG. A.,
van BoomJ. H.,
PlasterkR. H. A.1991b; Site-specific hydrolysis and alcoholysis of human immunodeficiency virus DNA termini mediated by the viral integrase protein. Nucleic Acids Research 19:6691–6698
VinkC.,
Oude GroenegerA. A. M.,
PlasterkR. H. A.1993; Identification of the catalytic and DNA-binding region of the human immunodeficiency virus type I integrase protein. Nucleic Acids Research 21:1419–1425
VinkC.,
van der LindenK. H.,
PlasterkR. H. A.1994; Activities of the feline immunodeficiency virus integrase protein produced in Escherichia coli. Journal of Virology 68:1468–1474
WoernerA. M.,
KlutchM.,
LevinJ. G.,
Marcus-SekuraC. J.1992; Localization of DNA binding activity of HIV-1 integrase to the C-terminal half of the protein. AIDS Research and Human Retroviruses 8:297–304
Comparative studies of bacterially expressed integrase proteins of caprine arthritis-encephalitis virus, maedi-visna virus and human immunodeficiency virus type 1