PMID- 10556302
OWN - NLM
STAT- MEDLINE
DCOM- 20000124
LR  - 20190513
IS  - 0964-6906 (Print)
IS  - 0964-6906 (Linking)
VI  - 8
IP  - 13
DP  - 1999 Dec
TI  - Characterization of SURF-1 expression and Surf-1p function in normal and disease 
      conditions.
PG  - 2533-40
AB  - Loss-of-function mutations of the SURF-1 gene have been associated with Leigh
      syndrome with cytochrome c oxidase (COX) deficiency. Mature Surf-1 protein
      (Surf-1p) is a 30 kDa hydrophobic polypeptide whose function is still unknown.
      Using antibodies against a recombinant, hemagglutinin-tagged Surf-1p, we have
      demonstrated that this protein is imported into mitochondria as a larger
      precursor, which is then processed into the mature product by cleaving off an
      N-terminal leader polypeptide of approximately 40 amino acids. By using western
      blot analysis with specific antibodies, we showed that Surf-1p is localized in
      and tightly bound to the mitochondrial inner membrane. The same analysis revealed
      that no protein is present in cell lines harboring loss-of-function mutations of 
      SURF-1, regardless of their type and position. Northern blot analysis showed the 
      virtual absence of specific SURF-1 transcripts in different mutant cell lines.
      This result suggests that several mutations of SURF-1 are associated with severe 
      mRNA instability. To understand better whether and which domains of the protein
      are essential for function, we generated several constructs with truncated or
      partially deleted SURF-1 cDNAs. None of these constructs, expressed into Surf-1p 
      null mutant cells, were able to rescue the COX phenotype, suggesting that
      different regions of the protein are all essential for function. Finally,
      experiments based on blue native two-dimensional gel electrophoresis indicated
      that assembly of COX in Surf-1p null mutants is blocked at an early step, most
      likely before the incorporation of subunit II in the nascent intermediates
      composed of subunit I alone or subunit I plus subunit IV. However, detection of
      residual amounts of fully assembled complex suggests a certain degree of
      redundancy of this system.
FAU - Tiranti, V
AU  - Tiranti V
AD  - Istituto Nazionale Neurologico 'C. Besta', Via Celoria 11, 20133 Milano, Italy.
FAU - Galimberti, C
AU  - Galimberti C
FAU - Nijtmans, L
AU  - Nijtmans L
FAU - Bovolenta, S
AU  - Bovolenta S
FAU - Perini, M P
AU  - Perini MP
FAU - Zeviani, M
AU  - Zeviani M
LA  - eng
GR  - 1180/Telethon/Italy
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Hum Mol Genet
JT  - Human molecular genetics
JID - 9208958
RN  - 0 (DNA, Mitochondrial)
RN  - 0 (Membrane Proteins)
RN  - 0 (Mitochondrial Proteins)
RN  - 0 (Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (SHY1 protein, S cerevisiae)
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - 0 (Surf-1 protein)
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - COS Cells
MH  - DNA, Mitochondrial/genetics
MH  - Electron Transport Complex IV/metabolism
MH  - Electrophoresis, Gel, Two-Dimensional
MH  - Genetic Complementation Test
MH  - Humans
MH  - Leigh Disease/enzymology/genetics/*metabolism
MH  - Membrane Proteins/genetics/metabolism
MH  - Mitochondria/metabolism
MH  - Mitochondrial Proteins
MH  - Mutagenesis, Site-Directed
MH  - Mutation
MH  - Proteins/genetics/*metabolism
MH  - Recombinant Proteins/genetics/metabolism
MH  - *Saccharomyces cerevisiae Proteins
EDAT- 1999/11/11 00:00
MHDA- 1999/11/11 00:01
CRDT- 1999/11/11 00:00
PHST- 1999/11/11 00:00 [pubmed]
PHST- 1999/11/11 00:01 [medline]
PHST- 1999/11/11 00:00 [entrez]
AID - ddc286 [pii]
AID - 10.1093/hmg/8.13.2533 [doi]
PST - ppublish
SO  - Hum Mol Genet. 1999 Dec;8(13):2533-40. doi: 10.1093/hmg/8.13.2533.