PMID- 24124581
OWN - NLM
STAT- MEDLINE
DCOM- 20140522
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 10
DP  - 2013
TI  - Biochemical characterization of novel retroviral integrase proteins.
PG  - e76638
LID - 10.1371/journal.pone.0076638 [doi]
LID - e76638
AB  - Integrase is an essential retroviral enzyme, catalyzing the stable integration of 
      reverse transcribed DNA into cellular DNA. Several aspects of the integration 
      mechanism, including the length of host DNA sequence duplication flanking the 
      integrated provirus, which can be from 4 to 6 bp, and the nucleotide preferences 
      at the site of integration, are thought to cluster among the different retroviral 
      genera. To date only the spumavirus prototype foamy virus integrase has provided 
      diffractable crystals of integrase-DNA complexes, revealing unprecedented details 
      on the molecular mechanisms of DNA integration. Here, we characterize five 
      previously unstudied integrase proteins, including those derived from the 
      alpharetrovirus lymphoproliferative disease virus (LPDV), betaretroviruses 
      Jaagsiekte sheep retrovirus (JSRV), and mouse mammary tumor virus (MMTV), 
      epsilonretrovirus walleye dermal sarcoma virus (WDSV), and gammaretrovirus 
      reticuloendotheliosis virus strain A (Rev-A) to identify potential novel 
      structural biology candidates. Integrase expressed in bacterial cells was 
      analyzed for solubility, stability during purification, and, once purified, 3' 
      processing and DNA strand transfer activities in vitro. We show that while we 
      were unable to extract or purify accountable amounts of WDSV, JRSV, or LPDV 
      integrase, purified MMTV and Rev-A integrase each preferentially support the 
      concerted integration of two viral DNA ends into target DNA. The sequencing of 
      concerted Rev-A integration products indicates high fidelity cleavage of target 
      DNA strands separated by 5 bp during integration, which contrasts with the 4 bp 
      duplication generated by a separate gammaretrovirus, the Moloney murine leukemia 
      virus (MLV). By comparing Rev-A in vitro integration sites to those generated by 
      MLV in cells, we concordantly conclude that the spacing of target DNA cleavage is 
      more evolutionarily flexible than are the target DNA base contacts made by 
      integrase during integration. Given their desirable concerted DNA integration 
      profiles, Rev-A and MMTV integrase proteins have been earmarked for structural 
      biology studies.
FAU - Ballandras-Colas, Allison
AU  - Ballandras-Colas A
AD  - Department of Cancer Immunology & AIDS, Dana-Farber Cancer Institute, and 
      Department of Medicine, Harvard Medical School, Boston, Massachusetts, United 
      States of America.
FAU - Naraharisetty, Hema
AU  - Naraharisetty H
FAU - Li, Xiang
AU  - Li X
FAU - Serrao, Erik
AU  - Serrao E
FAU - Engelman, Alan
AU  - Engelman A
LA  - eng
GR  - R01 AI070042/AI/NIAID NIH HHS/United States
GR  - T32 AI007245/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20131004
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (DNA, Viral)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Viral Proteins)
RN  - EC 2.7.7.- (Integrases)
SB  - IM
MH  - Animals
MH  - DNA, Viral/genetics/metabolism
MH  - Gene Expression
MH  - Humans
MH  - Integrases/*chemistry/genetics/isolation & purification/metabolism
MH  - Proviruses/genetics
MH  - Recombinant Proteins/chemistry/genetics/isolation & purification/metabolism
MH  - Retroviridae/*enzymology/genetics
MH  - Solubility
MH  - Viral Proteins/*chemistry/genetics/isolation & purification/metabolism
MH  - Virus Integration/physiology
PMC - PMC3790719
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2013/10/15 06:00
MHDA- 2014/05/23 06:00
PMCR- 2013/10/04
CRDT- 2013/10/15 06:00
PHST- 2013/07/29 00:00 [received]
PHST- 2013/08/12 00:00 [accepted]
PHST- 2013/10/15 06:00 [entrez]
PHST- 2013/10/15 06:00 [pubmed]
PHST- 2014/05/23 06:00 [medline]
PHST- 2013/10/04 00:00 [pmc-release]
AID - PONE-D-13-30616 [pii]
AID - 10.1371/journal.pone.0076638 [doi]
PST - epublish
SO  - PLoS One. 2013 Oct 4;8(10):e76638. doi: 10.1371/journal.pone.0076638. eCollection 
      2013.