3-Hydroxy-3-methylglutaryl-coenzyme-A synthase from Blattella germanica. Cloning, expression, developmental pattern and tissue expression.
Insects do not synthesize cholesterol; the 3-hydroxy-3-methylglutaryl-coenzyme-A (HMG-CoA) produced by HMG-CoA synthase is transformed to mevalonate by HMG-CoA reductase for the production of non-sterol isoprenoids, which are essential for growth and differentiation. To understand the regulation and developmental role of HMG-CoA synthase, we have cloned a 1658 bp cDNA that encompasses the entire transcription unit of the HMG-CoA synthase gene from the cockroach Blattella germanica. This cDNA clone was isolated using as a probe a partial cDNA of B. germanica HMG-CoA synthase, amplified using the polymerase chain reaction. Analysis of the nucleotide sequence reveals that the cDNA encodes a polypeptide of 453 amino acids (M(r) 50338) that is similar to vertebrate HMG-CoA synthase (74-76% conserved residues). The B. germanica cDNA has been expressed as a fusion protein in Escherichia coli and exhibits HMG-CoA synthase activity. The HMG-CoA synthase transcript was differentially expressed throughout B. germanica development. Analysis of RNA samples from different adult female tissues shows high HMG-CoA synthase mRNA levels in the ovary and lower levels in brain and muscle.
Martínez-González
J
J
Unidad de Bioquímica, Facultad de Farmacia, Universidad de Barcelona, Spain.
Buesa
C
C
Piulachs
M D
MD
Bellés
X
X
Hegardt
F G
FG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003058:Cockroaches / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D016415:Sequence Alignment; D014158:Transcription, Genetic
D018076:DNA, Complementary; D006904:Hydroxymethylglutaryl-CoA Synthase