The structure of the human NDUFV1 gene encoding the 51-kDa
subunit of mitochondrial complex I
Rene´ F.M. de Coo,
Paul A.L. Buddiger,
Hubert J.M. Smeets,
* Bernard A. van Oost
Dept. of Human Genetics, University Hospital Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands
Dept. of Child Neurology, University Hospital Rotterdam, P.O. Box 2060, 3000 CB Rotterdam, The Netherlands
Received: 5 May 1998 / Accepted: 28 August 1998
Abstract. The genomic organization of the human 51-kDa sub-
unit gene (NDUFV1) on human Chromosome (Chr) 11q13 was
determined. The NDUFV1 gene consists of 10 exons. Exon 1
encodes for the 20-amino-acids-long import sequence, and exon 1
through 10 codes for the 444-amino-acids-long mature protein.
The protein sequence is highly conserved between human and
bovine. Northern blotting analysis showed that the NDUFV1 gene
expression varies widely among tissues and that in testis a unique
mRNA species is present. In comparison with the other complex I
flavoproteins, the expression of the 51-kDa gene in pancreatic
tissue is high.
NADH:ubiquinone oxidoreductase (complex I) of the mitochon-
drial respiratory chain consists of at least 41 subunits, of which all
but seven are nuclearly encoded. Complex I removes electrons
from NADH and passes them on to the electron acceptor ubiqui-
none. This is the first step in the process of electron transfer
through a number of intermediates to oxygen and drives the pro-
duction of ATP (Walker 1992). With chaotropic reagents, complex
I can be fragmented into three fractions (Ragan 1987): two hydro-
philic fractions, the flavoprotein and the iron protein fraction, and
one hydrophobic fraction. The flavoprotein fraction contains three
subunits of 51 kDa, 24 kDa, and 10 kDa, and the oxidation of
NADH is mediated by this fraction. The 51 kDa is directly in-
volved in the transfer of two electrons from NADH to ubiquinone,
with the simultaneous translocation of four electrons across the
mitochondrial inner membrane (Deng et al. 1990). The 51-kDa
protein probably contains binding sites for both the cofactor and
for the oxidant of NADH, FMN (Patel et al. 1991; Krishnamoorthy
and Hinkle 1988), as well as a binding site for a tetranuclear
iron-sulfur center (Ohnishi et al. 1985). The 51-kDa protein is an
evolutionary conserved subunit since there is a strong amino acid
sequence homology with part of the bacterial hydrogenase gene of
Alcaligenes eutrophus. The gene for the ␣-subunit of the hydrog-
enase resides in the hoxF (␣) part of the hoxS locus in Alcaligenes
eutrophus (Tran-Betcke et al. 1990) and was shown to encode a
fusion of closely related sequences from the 51- (Pilkington et al.
1991) and 24-kDa (Pilkington and Walker, 1989) subunits. The
same relationships were reported in Neurospora crassa (Preis et al.
1991) and Paracoccus denitrificans (Xu et al. 1991).
In 1991 the bovine cDNA for the 51-kDa subunit was cloned
by Patel (Patel et al. 1991) and Pilkington (Pilkington et al. 1991).
By serendipity, Spencer (Spencer et al. 1992) localized the
NDUFV1 gene to be 30,000 bp downstream of the GSTP1 gene on
a cosmid originating from human Chr 11. In 1993 Ali and asso-
ciates sequenced the partial human 51-kDa subunit cDNA and
used this cDNA to confirm the chromosomal localization on 11q13
*Present address: Division of Genetics, University of Limburg, P.O. Box
616, 6200 MD Maastricht, The Netherlands
**Present address: Dept. of Clinical Sciences of Companion Animals,
University of Utrecht, P.O. Box 80154, 3508 TD Utrecht, The Netherlands
Part of the results were presented at the annual meeting of the American
Society of Human Genetics held on October 28 - November 1 1997 in
Baltimore, Maryland: P.A.L. Buddiger, W. Ruitenbeek, H.R. Scholte, B.A.
van Oost, H.J.M. Smeets, I.F.M. de Coo. Molecular genetic analysis of
complex I genes in patients with a deficiency of the respiratory chain. Am
J Hum Genet, 61, A305, 1997.
The nucleotide sequence data reported in this paper have been submitted to
EMBL and have been assigned the accession number Y16309 (Y17379-
17383) and Y16310.
Correspondence to: R.F.M. de Coo at Dept. of Child Neurology.
Table 1. Oligonucleotides used for PCR and cycle sequencing of the NDUFV1 gene.
Exon Sequences (5Ј–3Ј)
1128f −21– −1 intron 1 CCTGACCCTTTGTCTCCCTAG
1078f 456–476 IV CGCTGCCTATATCTACATCCG
1112r +45– +22 intron 4 GCACACTGAACACCTTCTCATCTG
1081f 605–625 V GGGCCTACATCTGTGGAGAGG
1083r 644–625 V GACTCGATGAGCGCTGTCTC
1117f 865–885 VI GAGGAGGAGATGTCTGTGCCC
1080r 885–865 VI GGGCACAGACATCTCCTCCTC
1143f 934–955 VII GACTACCTCCTTGCTGTGATCC
1150r 1163–1145 VIII CACCGGTCACATGGGGTAC
1181f 1136–1154 VIII GTGGCCAGTGTACCCCATG
1147r 1430–1410 X TGGATAGACGCAGGACAGCAG
f, forward primer; r, reverse primer.
Nucleotide numbering according to cDNA sequence, Fig. 3.
Fig. 1. Hybridization of the cDNA for
the bovine 51-kDa protein with
restriction digests of human genomic
DNA. This probe contains the entire
coding region. The human DNA was
digested with the following restriction
enzymes: 1, PvuII; 2, StuI; 3, SstI; 4,
HindIII; 5, EcoRI. The size marker used
is the 1-kb ladder from Gibco BRL.
Mammalian Genome 10, 49–53 (1999).
© Springer-Verlag New York Inc. 1999