The KlPGS1 gene encoding phosphatidylglycerolphosphate synthase in Kluyveromyces lactis is essential and assigned to chromosome I.
The phosphatidylglycerolphosphate synthase (CDP-diacylglycerol:sn-glycerol-3-phosphate 3-phosphatidyltransferase, EC 2.7.8.5) is an essential enzyme in biosynthesis of cardiolipin. In this work we report the isolation, heterological cloning, molecular characterization and physical mapping of the Saccharomyces cerevisiae PEL1/PGS1 homologue from Kluyveromyces lactis. The pel1 mutant strain of S. cerevisiae was used to isolate this homologue by screening a K. lactis genomic library. The novel cloned gene was named KlPGS1. Its coding region was found to consist of 1623 bp. The corresponding protein exhibits 55% amino acid identity to its S. cerevisiae counterpart. The presence of the mitochondrial presequence indicates its mitochondrial localization. Sporulation and ascus dissection of diploids heterozygous for single-copy disruption of KlPGS1 revealed that the KlPGS1 gene, is essential in K. lactis. Using a DIG-dUTP-labeled DNA probe-originated from the KlPGS1 gene and Southern hybridization of contour-clamped homogeneous electric field (CHEF)-separated K. lactis chromosomal DNA, the KlPGS1 gene was assigned to chromosome I. The nucleotide sequence data reported in this paper were submitted to GenBank and assigned the Accession No. AY176328.
Tyciakova
Silvia
S
Department of Microbiology and Virology, Faculty of Sciences, Comenius University in Bratislava, Mlynska dolina B-2, 842 15 Bratislava, Slovak Republic.
Obernauerova
Margita
M
Dokusova
Lucia
L
Kooistra
Rolf A
RA
Steensma
H Yde
HY
Sulo
Pavol
P
Subik
Julius
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase / Q000235:genetics*; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal / Q000235:genetics*; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D005800:Genes, Fungal / Q000235:genetics*; D007716:Kluyveromyces / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment
D004271:DNA, Fungal; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase