PMID- 15547949
OWN - NLM
STAT- MEDLINE
DCOM- 20050715
LR  - 20131121
IS  - 0730-2312 (Print)
IS  - 0730-2312 (Linking)
VI  - 94
IP  - 4
DP  - 2005 Mar 1
TI  - Evidence of a dominant negative mutant of yeast methionine aminopeptidase type 2 
      in Saccharomyces cerevisiae.
PG  - 656-68
AB  - Eukaryotic methionine aminopeptidase type 2 (MetAP2, MetAP2 gene (MAP2)), 
      together with eukaryotic MetAP1, cotranslationally hydrolyzes initiator 
      methionine from nascent polypeptides when the side chain of the second residue is 
      small and uncharged. In this report, we took advantage of the yeast 
      (Saccharomyces cerevisiae) map1 null strain's reliance on MetAP2 activity for the 
      growth and viability to provide evidence of the first dominant negative mutant of 
      eukaryotic MetAP2. Replacement of the conserved His(174) with alanine within the 
      C-terminal catalytic domain of yeast MetAP2 eliminated detectable catalytic 
      activity against a peptide substrate in vitro. Overexpression of MetAP2 (H174A) 
      under the strong GPD promoter in a yeast map1 null strain was lethal, whereas 
      overexpression under the weaker GAL1 promoter slightly inhibited map1 null 
      growth. Deletion mutants further revealed that the N-terminal region of MetAP2 
      (residues 2-57) is essential but not sufficient for MetAP2 (H174A) to fully 
      interfere with map1 null growth. Together, these results indicate that 
      catalytically inactive MetAP2 is a dominant negative mutant that requires its 
      N-terminal region to interfere with wild-type MetAP2 function.
FAU - Vetro, Joseph A
AU  - Vetro JA
AD  - Edward A. Doisy Department of Biochemistry and Molecular Biology, St. Louis 
      University Health Sciences Center, 1402 S. Grand Blvd., St. Louis, MO 63104, USA.
FAU - Dummitt, Benjamin
AU  - Dummitt B
FAU - Micka, William S
AU  - Micka WS
FAU - Chang, Yie-Hwa
AU  - Chang YH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - J Cell Biochem
JT  - Journal of cellular biochemistry
JID - 8205768
RN  - 4QD397987E (Histidine)
RN  - EC 3.4.11.- (Aminopeptidases)
RN  - EC 3.4.11.18 (methionine aminopeptidase 2)
RN  - EC 3.4.24.- (Metalloendopeptidases)
SB  - IM
MH  - Aminopeptidases/*genetics/*metabolism
MH  - Catalysis
MH  - Cell Survival/genetics
MH  - Gene Expression Regulation, Fungal
MH  - Genes, Dominant/genetics
MH  - Histidine/genetics/metabolism
MH  - Metalloendopeptidases/*genetics/*metabolism
MH  - Mutation/*genetics
MH  - Promoter Regions, Genetic/genetics
MH  - Saccharomyces cerevisiae/cytology/*enzymology/*genetics
EDAT- 2004/11/18 09:00
MHDA- 2005/07/16 09:00
CRDT- 2004/11/18 09:00
PHST- 2004/11/18 09:00 [pubmed]
PHST- 2005/07/16 09:00 [medline]
PHST- 2004/11/18 09:00 [entrez]
AID - 10.1002/jcb.20285 [doi]
PST - ppublish
SO  - J Cell Biochem. 2005 Mar 1;94(4):656-68. doi: 10.1002/jcb.20285.