Document 0636
 DOCN  M9550636
 TI    Characterization of the human spuma retrovirus integrase by
       site-directed mutagenesis, by complementation analysis, and by swapping
       the zinc finger domain of HIV-1.
 DT    9505
 AU    Pahl A; Flugel RM; Abteilung Retrovirale Genexpression, Deutsches;
       Krebsforschungszentrum, Heidelberg, Federal Republic of Germany.
 SO    J Biol Chem. 1995 Feb 17;270(7):2957-66. Unique Identifier : AIDSLINE
       MED/95155376
 AB    The human spuma retrovirus or foamy virus integrase (HFV IN) is an
       enzymatically active protein consisting of domains similar to other
       retroviral integrases: an amino-terminal HH-CC finger, a centrally
       located region with the conserved D, D-35-E protein motif required for
       catalytic activity and oligomerization, and at least one DNA binding
       domain implicated in the 3' DNA processing activity and integrase.
       Recombinant, purified HFV IN protein carrying 10 histidine residues
       displays a site-specific endonuclease, an integrase, and a disintegrase
       activity with oligonucleotide substrates that mimic the viral long
       terminal repeat (LTR) ends. Site-directed mutagenesis of conserved HFV
       IN residues of the catalytic domain had increased endonuclease and
       disintegrase activities. Deletion mutants at both ends of the HFV IN
       protein were generated, purified, and characterized. Unexpectedly, it
       was found that the HFV integrase and disintegrase activities require an
       intact NH2-terminal sequence and that COOH-terminal deletions led to an
       increase in disintegrase activity. The HH-CC finger of HFV IN was
       exchanged with that of the human immunodeficiency virus-1 (HIV-1) IN
       protein. The resulting chimeric IN had a 3' processing activity that
       utilized the HFV LTR instead of the HIV LTR, indicating that the central
       domain is crucial for substrate recognition. Functional complementation
       of the amino-terminal deletion mutant of HFV IN was achieved by a
       carboxyl-terminal deletion mutant of the chimeric IN, resulting in high
       levels of integrase activity.
 DE    Amino Acid Sequence  Base Sequence  Chimeric Proteins/METABOLISM
       Comparative Study  DNA Nucleotidyltransferases/CHEMISTRY/ISOLATION &
       PURIF/  *METABOLISM  DNA Primers  Genetic Complementation Test  Human
       Hydrogen-Ion Concentration  HIV Long Terminal Repeat  HIV-1/*ENZYMOLOGY
       Kinetics  Molecular Sequence Data  Mutagenesis, Site-Directed
       Oligodeoxyribonucleotides/CHEMISTRY/CHEMICAL SYNTHESIS/METABOLISM
       Recombinant Proteins/CHEMISTRY/ISOLATION & PURIF/METABOLISM  *Repetitive
       Sequences, Nucleic Acid  Retroviridae/*ENZYMOLOGY  Sequence Deletion
       Spumavirus/*ENZYMOLOGY  Substrate Specificity  *Zinc Fingers  JOURNAL
       ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
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