Glycerol 3-phosphate dehydrogenase gene expression in cultured 3T3-L1 adipocytes: regulation by insulin, dexamethasone and dibutyryl cAMP at the level of mRNA abundance, transcription and mRNA stability.
In fully differentiated 3T3-L1 adipocytes, glycerol 3-phosphate dehydrogenase (G3PDH:Sn-glycerol 3-phosphate:NAD+ 2-oxidoreductase, EC 1.1.1.8) is subject to regulation by hormones and dibutyryl cAMP. An increase by insulin (4-fold) and decrease by dexamethasone (by 50%) and dibutyryl cAMP (by 70%) was observed for G3PDH mRNA abundance as analyzed by Northern blot hybridization. In addition, incubation of adipocytes with dibutyryl cAMP resulted in 3-fold increase in G3PDH gene transcription as measured by nuclear transcript elongation assay. The effects of these modulators on the control of G3PDH mRNA stability were also investigated. The G3PDH mRNA has a half-life of about 125 min. Dibutyryl cAMP caused an increase in G3PDH mRNA degradation by greater than 2-fold (t1/2 = 55 min) whereas insulin had an opposite effect (t1/2 = 240 min) and dexamethasone was without any effect on G3PDH mRNA stability. Taken together, our results directly demonstrate that in fully differentiated 3T3-L1 adipocytes the regulation of G3PDH gene expression by dibutyryl cAMP and insulin is exerted by alterations in transcription as well as mRNA stability.
Bhandari
B
B
Department of Medicine, University of Texas Health Science Center, San Antonio 78284.
Saini
K S
KS
Miller
R E
RE
D016475:3T3 Cells; D000273:Adipose Tissue / Q000166:cytology / Q000201:enzymology*; D000818:Animals; D015152:Blotting, Northern; D003994:Bucladesine / Q000494:pharmacology*; D003907:Dexamethasone / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic; D005993:Glycerolphosphate Dehydrogenase / Q000235:genetics*; D007328:Insulin / Q000502:physiology*; D051379:Mice; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic
D007328:Insulin; D012333:RNA, Messenger; D003994:Bucladesine; D003907:Dexamethasone; D005993:Glycerolphosphate Dehydrogenase