Androgen receptor immunoreactivity of male rat cervical motor neurons is increased by chronic pharmacologic testosterone treatment.

Chronic pharmacologic testosterone treatment of adult male rats decreases neuromuscular transmission failure (NTF) in the rat diaphragm muscle and increases choline acetyltransferase (ChAT) mRNA levels in cervical motor neurons. These testosterone-induced changes in NTF and ChAT mRNA levels may be mediated through the activation of the androgen receptor (AR). The purpose of this study was to determine if the AR expression of cervical motor neurons is modulated by chronic pharmacologic testosterone treatment of gonadally intact male rats with testosterone propionate (TP). Serum testosterone levels were elevated by a subcutaneous implant of capsules containing crystalline TP for 28 days. The proportion of motor neurons containing AR-I positive nuclei was increased from 14.8 +/- 10.8% among the control group to 81.7 +/- 12.6% in the TP-treated animals (p<0.05). These results imply that anabolic-androgenic steroid effects on neuromuscular function may be mediated through AR dependent regulation of gene expression in motor neurons.

Phylogenetic characterization of novel transport protein families revealed by genome analyses.

As a result of recent genome sequencing projects as well as detailed biochemical, molecular genetic and physiological experimentation on representative transport proteins, we have come to realize that all organisms possess an extensive but limited array of transport protein types that allow the uptake of nutrients and excretion of toxic substances. These proteins fall into phylogenetic families that presumably reflect their evolutionary histories. Some of these families are restricted to a single phylogenetic group of organisms and may have arisen recently in evolutionary time while others are found ubiquitously and may be ancient. In this study we conduct systematic phylogenetic analyses of 26 families of transport systems that either had not been characterized previously or were in need of updating. Among the families analyzed are some that are bacterial-specific, others that are eukaryotic-specific, and others that are ubiquitous. They can function by either a channel-type or a carrier-type mechanism, and in the latter case, they are frequently energized by coupling solute transport to the flux of an ion down its electrochemical gradient. We tabulate the currently sequenced members of the 26 families analyzed, describe the properties of these families, and present partial multiple alignments, signature sequences and phylogenetic trees for them all.

The LysE superfamily: topology of the lysine exporter LysE of Corynebacterium glutamicum, a paradyme for a novel superfamily of transmembrane solute translocators.

In Corynebacterium glutamicum the LysE carrier protein exhibits the unique function of exporting L-lysine. We here analyze the membrane topology of LysE, a protein of 236 amino acyl residues, using PhoA- and LacZ-fusions. The amino-terminal end of LysE is located in the cytoplasm whereas the carboxy-terminal end is found in the periplasm. Although 6 hydrophobic domains were identified based on hydropathy analyses, only five transmembrane spanning helices appear to be present. The additional hydrophobic segment may dip into the membrane or be surface localized. We show that LysE is a member of a family of proteins found, for example, in Escherichia coil, Bacillus subtilis, Mycobacterium tuberculosis and Helicobacter pylori. This family, which we have designated the LysE family, is distantly related to two additional protein families which we have designated the YahN and CadD families. These three families, the members of which exhibit similar sizes, hydropathy profiles, and sequence motifs comprise the LysE superfamily. Functionally characterized members of the LysE superfamily export L-lysine, cadmium and possibly quarternary amines. We suggest that LysE superfamily members will prove to catalyze export of a variety of biologically important solutes.

CHR, a novel family of prokaryotic proton motive force-driven transporters probably containing chromate/sulfate antiporters.

We describe a small family of proteins, CHR, which contains members that function in chromate and/or sulfate transport. CHR proteins occur in bacteria and archaea. They consist of about 400 amino acyl residues, appear to have 10 transmembrane alpha-helical segments in an unusual 4+6 arrangement, and arose by an intragenic duplication event.

Plasmid-determined cadmium resistance in Pseudomonas putida GAM-1 isolated from soil.

Cadmium-resistant Pseudomonas putida GAM-1, which was able to grow in concentrations of CdCl2 as high as 7 mM, was isolated from soil in a rice paddy. This bacterium harbored a DNA plasmid of about 52 kilobases. The plasmid (pGU100) transformed Escherichia coli C600 to cadmium resistance. A cadmium-resistant transformant of E. coli C600 contained a plasmid corresponding to that seen in P. putida GAM-1. The transformant did not take up cadmium as well as P. putida GAM-1 did.