Uncoupling proteins (UCPs) are mitochondrial membrane proton transporters that uncouple oxidative phosphorylation by dissipating the proton gradient across the membrane. We have investigated regulation of the UCP3 gene in skeletal muscle and C2C12 muscle cells. UCP3 mRNA in mouse skeletal muscle is markedly increased by fasting and rapidly (within 4 h) decreased by re-feeding. Methyl palmoxirate, which inhibits fatty acid uptake by mitochondria and increases blood free fatty acids, prevents the fall in UCP3 message level induced by re-feeding. These findings suggest that fatty acid or a metabolite thereof, activates the UCP3 gene. Proof that fatty acid per se up-regulates UCP3 mRNA was obtained with C2C12 muscle cells in culture. Thus, oleic acid activated expression of UCP3 mRNA in differentiated C2C12 myotubes in a time and concentration-dependent manner. Moreover, BRL49653, a ligand for the nuclear hormone receptor PPARgamma induces expression of UCP3 mRNA suggesting that PPARgamma may regulate transcription of the UCP3 gene.

Adipose tissue has long been known to house the largest energy reserves in the animal body. Recent research indicates that in addition to this role, the adipocyte functions as a global regulator of energy metabolism. Adipose tissue is exquisitely sensitive to a variety of endocrine and paracrine signals, e.g. insulin, glucagon, glucocorticoids, and tumor necrosis factor (TNF), that combine to control both the secretion of other regulatory factors and the recruitment and differentiation of new adipocytes. The process of adipocyte differentiation is controlled by a cascade of transcription factors, most notably those of the C/EBP and PPAR families, which combine to regulate each other and to control the expression of adipocyte-specific genes. One such gene, i.e. the obese gene, was recently identified and found to encode a hormone, referred to as leptin, that plays a major role in the regulation of energy intake and expenditure. The hormonal and transcriptional control of adipocyte differentiation is discussed, as is the role of leptin and other factors secreted by the adipocyte that participate in the regulation of adipose homeostasis.

Appressorium formation in germinating Colletotrichum gloeosporioides is induced by the surface wax of the host, the avocado fruit. To elucidate the mechanism by which differentiation of appressorium formation is induced, the fungal genes specifically activated by this host signal were sought. From a cDNA library of the transcripts present in appressorium-forming conidia, the clones representing nongerminating conidia were removed by hybridization with cDNAs synthesized from the nongerminating conidia. From this subtracted library, clones that hybridized with cDNA for transcripts from appressorium-forming conidia and not with cDNA for transcripts from germinating conidia were selected. Three such clones were isolated and sequenced. The genes for these three transcripts were also cloned and sequenced. Northern blot analysis showed that transcripts that hybridized with these three clones were expressed in the conidium only during the process of appressorium formation induced by avocado surface wax, and that these transcripts were not detectable when appressorium formation was prevented even in the presence of avocado wax. Nucleotide sequences of the clones revealed that one clone, cap3, contained an open reading frame (ORF) that would code for a 26-amino acid, cysteine-rich peptide with significant homology to Neurospora crassa copper metallothionein. Another clone, cap5, contained an ORF that would code for a 27-amino acid cysteine-rich peptide with less homology to metallothioneins. Cu2+ and Cd2+ also induced the expression of these genes at lower levels. The histochemical analysis of transformants containing the cap5 promoter fused to the beta-glucuronidase (GUS) gene showed that the cap5 gene promoter caused GUS expression exclusively during appressorium formation and most of the gus activity was in the appressorium. The cap22 clone contained an ORF coding for a 227-amino acid polypeptide of 22 kDa, which did not show significant homology to any known proteins. Recombinant CAP22 protein was produced using a pET-19b expression system in Escherichia coli, purified, and used to prepare rabbit antibodies. Western blot analysis of proteins from the appressorium-forming conidia revealed a major cross-reacting protein at 43 kDa and a minor band at 68 kDa, indicating that the potential glycosylation sites found in the primary translation product were probably glycosylated. Results of immunogold localization showed that CAP22 protein was located on the wall of the appressorium.