Characterization of the Functional Domains of Human Foamy Virus Integrase Using Chimeric Integrases

다국어 초록 (Multilingual Abstract)

Retroviral integrases insert viral DNA into target DNA. In this process they recognize their own DNA specifically via functional domains. In order to analyze these
functional domains, we constructed six chimeric integrases by swapping domains between HIV-1 and HFV integrases, and two point mutants of HFV integrase.
Chimeric integrases with the central domain of HIV-1 integrase had strand transfer and disintegration activities, in agreement with the idea that the central domain determines viral DNA specificity and has catalytic
activity. On the other hand, chimeric integrases with the central domain of HFV integrase did not have any enzymatic activity apart from FFH that had weak disintegration activity, suggesting that the central domain
of HFV integrase was defective catalytically or structurally. However, these inactive chimeras were efficiently complemented by the point mutants (D164A and E200A) of HFV integrase, indicating that the central
domain of HFV integrase possesses potential enzymatic activity but is not able to recognize viral or target DNA without the help of its homologous Nterminal and C-terminal domains.

참고문헌 (Reference)

1 "structure of the initial covalentproduct and its precursor and a role for the viral IN protein" -86, 1989

2 "structure of an integration intermediate" 1988

3 "mechanism of viral DNA cleavage and DNAstrand transfer" 1991

4 "anovel endonuclease activity of human immunodeficiency virustype 1 and other retroviral integrases" (n-specific alcoholysis) : 1996

5 "The coreand carboxylterminal domains of the integrase protein ofhuman immunodeficiency virus type 1 each contribute tononspecific DNA binding" 1994

6 "The avian retroviral integration protein cleaves the terminalsequences of linear viral DNA at the in vivo sites of integration" 1989

7 "The N-terminal region ofHIV-1 integrase is required for integration but for DNAbinding" 1992

8 "Stereospecificityof reactions catalyzed by HIV-1 integrase" 1999

9 "Solution structure of the DNA bindingdomain of HIV-1 integrase" 1995

10 "Site directedmutagenesis of HIV-1 integrase demonstrates differential effectson integrase functions in vitro" 1993

11 "Roleof the nonspecific DNA-binding region and ？ helices withinthe core domain of retroviral integrase in selecting targetDNA sites for integration" 2001

12 "Reversal of integration and DNA splicing mediatedby integrase of human immunodeficiency virus" Vincent -255, 1992

13 "Residues critical for retroviral integrativerecombination in a region that is highly conservedamong retroviral/retrotransposon integrases and bacterial insertionsequence transposases" 1992

14 "Polynucleotidyl transfer reaction in transpositoinalDNA recombination" 1992

15 "Nucleotide sequence analysis of the env gene and its flankingregions of the human spumaretrovirus reveals two novelgenes" 1987

16 "Novel and recurrent mutations of the LDL receptorgene in Korean patients with familial hypercholesterolemia" -18, 2004

17 "Mapping viral DNA specificityto the central region of integrase by using functionalhuman immunodeficiency virus type 1/visna virus chimericprotein" 1998

18 "Mapping target site selection for the non-specificnuclease activities of retroviral integrase" puf (puf): 2000

19 "Mapping domains of retroviralintegrase responsible for viral DNA specificity and targetsite selection by analysis of chimeras between human immunodeficiencyvirus type 1 and visna virus integrase" 1995

20 "Localization of DNA binding activity of HIV-1 integraseto the C-terminal half of the protein" -8, 1992

21 "Intramolecularchimeras of the p51 subunit between HIV-1 and FIV reversetranscriptase suggest a stabilizing function for the p66subunit in the heterodimeric enzyme" 1999

22 "Identificationof discrete functional domains of HIV-1 integrase andtheir organization within an active multimeric complex" 1993

23 "Identification of the catalytic and DNA-binding region of thehuman immunodeficiency virus type 1 integrase protein" 1993

24 "Identification of conservedamino acid residues critical for human immunodeficiency virustype 1 integrase function in vitro" 1992

25 "Genetic analysis ofhomomeric interaction of human immunodeficiency virustype 1 integrase using the yeast two-hybrid system" -90, 1993

26 "Genetic analysis of the human immunodeficiency virustype 1 integrase protein" 1994

27 "Gene splicing by overlap extension" 1993

28 "Endonucleolytic cleavagesand DNA-joining activities of the integration protein of humanfoamy virus" 1993

29 "Domains of the integrase protein of humanimmunodeficiency virus type 1 responsible for polynucleotidyltransfer and zinc binding" -90, 1993

30 "Dissectingthe role of the N-terminal domain of human immunodeficiencyvirus integrase by trans-complementation analysis" 1999

31 "DNA bindingand the recognition of LTR sequence" 1991

32 "Complementation between HIV integrase proteinsmutated in different domains" -3267, 1993

33 "Comparison of enzymatic activitiesof the HIV-1 and HFV integrases to their U5 LTRsubstrates" 1999

34 "Characterizationof the minimal DNA-binding domain of HIV integraseprotein" 1994

35 "Characterization of the minimal DNA-binding domain ofHIV integrase protein" 1994

36 "Characterization of the humanspuma retrovirus intgrase by site-directed nutadenesis and by swapping the zinc finger domainof HIV-1" 1995

37 "Characterization of human immunodeficiency virustype 1 integrase expressed in Escherichia coli and analysis ofvariants with amino-terminal mutation" Vincent 1993

38 "Characterization of feline human immunodeficiencyvirus type 1 integrase and analysis of functional domains" 1997

39 "Characterization of chimeric enzymes between caprinearthritis-encephalitis virus maedi-visna virus and human immunodeficiencyvirus type 1 integrases expressed in Escherichiacoli" 2001

40 "Central core domain ofretroviral integrase is responsible for target site selection" 1997

41 "Binding aspects of baicalein to HIV-integrase" -12, 2001

42 "An unusual virus from a human nasopharyngeal carcinoma" 1971

43 "An essential interaction between distinct domains of HIV-1integrase mediates assembly of active multimer" Vincent 1995

44 "A stable complex betweenintegrase and viral DNA ends mediates HIV integration in vitro" -91, 1994

45 "A role forDNA-PK in retroviral DNA integration" -284, 1999

46 "A chimeric Ty3/Moloney murine leukemia virus integraseprotein is activity in vivo" 1998