Molecular cloning, developmental pattern and tissue expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase of the cockroach Blattella germanica.
In insects, 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA) synthesizes mevalonate for the production of nonsterol isoprenoids, which are essential for growth and differentiation. To understand the regulation and developmental role of HMG-CoA reductase, we have cloned a full-length HMG-CoA reductase cDNA from the cockroach Blattella germanica. This cDNA clone was isolated using as a probe a partial cDNA of B. germanica HMG-CoA reductase, amplified using the polymerase chain reaction. The composite 3433-bp cDNA sequence contains an open reading frame encoding a polypeptide of 856 amino acids (Mr, 93165). The C-terminal region is more similar to hamster HMG-CoA reductase than is the Drosophila melanogaster enzyme (79% and 69% conserved residues, respectively), and the potential transmembrane domains at the N-terminal region are structurally conservative with both enzymes. The C-terminal region of the B. germanica protein has been expressed as a fusion protein in Escherichia coli and exhibits HMG-CoA reductase activity. Analysis of B. germanica HMG-CoA reductase mRNA levels, reveals a 3.6-kb transcript, that is overexpressed in 4-day-old embryos. Northern-blot analysis of RNA samples from different adult female tissues shows high HMG-CoA reductase mRNA levels in the ovary and lower levels in brain and muscle.
Martinez-Gonzalez
J
J
Unidad de Bioquímica, Facultad de Farmacia, Universidad de Barcelona, Spain.
Buesa
C
C
Piulachs
M D
MD
Belles
X
X
Hegardt
F G
FG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003058:Cockroaches / Q000201:enzymology*; D004247:DNA; D004926:Escherichia coli; D005260:Female; D006903:Hydroxymethylglutaryl CoA Reductases / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D012333:RNA, Messenger; D004247:DNA; D006903:Hydroxymethylglutaryl CoA Reductases