PMID- 10074426
OWN - NLM
STAT- MEDLINE
DCOM- 19990422
LR  - 20071114
IS  - 0960-9822 (Print)
IS  - 0960-9822 (Linking)
VI  - 9
IP  - 4
DP  - 1999 Feb 25
TI  - Identification of a new uracil-DNA glycosylase family by expression cloning using
      synthetic inhibitors.
PG  - 174-85
AB  - BACKGROUND: The cellular environment exposes DNA to a wide variety of endogenous 
      and exogenous reactive species that can damage DNA, thereby leading to genetic
      mutations. DNA glycosylases protect the integrity of the genome by catalyzing the
      first step in the base excision-repair of lesions in DNA. RESULTS: Here, we
      report a strategy to conduct genome-wide screening for expressed DNA
      glycosylases, based on their ability to bind to a library of four synthetic
      inhibitors that target the enzyme's active site. These inhibitors, used in
      conjunction with the in vitro expression cloning procedure, led to the
      identification of novel Xenopus and human proteins, xSMUG1 and hSMUG1,
      respectively, that efficiently excise uracil residues from DNA. Despite a lack of
      statistically significant overall sequence similarity to the two established
      classes of uracil-DNA glycosylases, the SMUG1 enzymes contain motifs that are
      hallmarks of a shared active-site structure and overall protein architecture. The
      unusual preference of SMUG1 for single-stranded rather than double-stranded DNA
      suggests a unique biological function in ridding the genome of uracil residues,
      which are potent endogenous mutagens. CONCLUSIONS: The 'proteomics' approach
      described here has led to the isolation of a new family of uracil-DNA
      glycosylases. The three classes of uracil-excising enzymes (SMUG1 being the most 
      recently discovered) represent a striking example of structural and functional
      conservation in the almost complete absence of sequence conservation.
FAU - Haushalter, K A
AU  - Haushalter KA
AD  - Department of Chemistry and Chemical Biology Harvard University Cambridge
      Massachusetts 02138 USA.
FAU - Todd Stukenberg, M W
AU  - Todd Stukenberg MW
FAU - Kirschner, M W
AU  - Kirschner MW
FAU - Verdine, G L
AU  - Verdine GL
LA  - eng
SI  - GENBANK/AF125181
SI  - GENBANK/AF125182
GR  - 5 R01 GM 26875/GM/NIGMS NIH HHS/United States
GR  - GM 51330/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Curr Biol
JT  - Current biology : CB
JID - 9107782
RN  - 0 (Recombinant Proteins)
RN  - 0 (Xenopus Proteins)
RN  - EC 3.2.2.- (DNA Glycosylases)
RN  - EC 3.2.2.- (N-Glycosyl Hydrolases)
RN  - EC 3.2.2.- (SMUG1 protein, Xenopus laevis)
RN  - EC 3.2.2.- (SMUG1 protein, human)
RN  - EC 3.2.2.- (Uracil-DNA Glycosidase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Binding Sites
MH  - Cloning, Molecular
MH  - Conserved Sequence
MH  - *DNA Glycosylases
MH  - Humans
MH  - Molecular Sequence Data
MH  - N-Glycosyl Hydrolases/chemistry/*genetics/*metabolism
MH  - Protein Conformation
MH  - Recombinant Proteins/biosynthesis/chemistry/metabolism
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
MH  - Uracil-DNA Glycosidase
MH  - Xenopus
MH  - Xenopus Proteins
EDAT- 1999/03/13 00:00
MHDA- 1999/03/13 00:01
CRDT- 1999/03/13 00:00
PHST- 1999/03/13 00:00 [pubmed]
PHST- 1999/03/13 00:01 [medline]
PHST- 1999/03/13 00:00 [entrez]
AID - S0960-9822(99)80087-6 [pii]
PST - ppublish
SO  - Curr Biol. 1999 Feb 25;9(4):174-85.