Sequence and tissue-dependent RNA expression of mouse FAD-linked glycerol-3-phosphate dehydrogenase.
A 2432-bp cDNA for mouse FAD-linked glycerol-3-phosphate dehydrogenase, a nuclear-encoded enzyme associated with the inner mitochondrial membrane, was isolated from a Lambda ZAP phage library generated from brown adipocyte mRNA. The amino acid sequence was 95 and 93% homologous to the rat and human enzymes. PvuII and SacI polymorphisms between Mus spretus and C57BL/6J were used to map the mouse FAD-linked glycerol-3-phosphate dehydrogenase gene (Gdm1) to chromosome 2, 33 cM from the centromere. Northern blot analysis showed that brown adipose tissue predominantly expressed a 6.5-kb mRNA with lower expression of 4.5- and 2.4-kb forms, whereas brain and pancreatic islets almost exclusively expressed a 6.5-kb transcript, muscle expressed a 4.5-kb transcript, and testis expressed a 2.4-kb RNA form. Analysis of poly(A)+ RNA from brown adipose tissue suggested that all RNA forms are polyadenylated. Among the tissues examined, FAD-linked glycerol-3-phosphate dehydrogenase protein levels and enzyme activity were highest in brown adipose tissue, and a consistent correlation between protein levels and enzyme activity in all tissues was observed. RNA levels corresponded to protein levels in all tissues except testis, where high levels of 2.4-kb mRNA and relatively low protein were expressed. Exposing mice to cold temperatures induced the 6.5-kb mRNA and enzyme activity only in brown adipose tissue, suggesting a role for thermogenesis in this tissue. Although the molecular basis for the formation of the 6.5- and 4.5-kb mRNA's is not known, data suggest that the regulation of FAD-linked glycerol-3-phosphate dehydrogenase is complex and tissue-specific.
Koza
R A
RA
The Jackson Laboratory, Bar Harbor, Maine 04609, USA.
Kozak
U C
UC
Brown
L J
LJ
Leiter
E H
EH
MacDonald
M J
MJ
Kozak
L P
LP
D002001:Adipose Tissue, Brown / Q000201:enzymology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003080:Cold Temperature; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D005993:Glycerolphosphate Dehydrogenase / Q000235:genetics*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D014018:Tissue Distribution
D005182:Flavin-Adenine Dinucleotide; D005993:Glycerolphosphate Dehydrogenase