Enterococcus hirae (formerly Streptococcusfaecalis) ATCC 9790 has an F1F0-ATPase which functions as a regulator of the cytoplasmic pH but does not synthesize ATP. We isolated four clones which contained genes for c, b, delta, and alpha subunits of this enzyme but not for other subunit genes. It was revealed that two specific regions (upstream of the c-subunit gene and downstream of the gamma-subunit gene) were lost at a specific site in the clones we isolated, suggesting that these regions were unstable in Escherichia coli. The deleted regions were amplified by polymerase chain reaction, and the nucleotide sequences of these regions were determined. The results showed that eight genes for a, c, b, delta, alpha, gamma, beta, and epsilon subunits were present in this order. Northern (RNA) blot analysis showed that these eight genes were transcribed to one mRNA. The i gene was not found in the upper region of the a-subunit gene. Instead of the i gene, this operon contained a long untranslated region (240 bp) whose G+C content was only 30%1. There was no typical promoter sequence such as was proposed for E. coli, suggesting that the promoter structure of this species is different from that of E. coli. Deduced amino acid sequences suggested that E. hirae H+-ATPase is a typical F1F0-type ATPase but that its gene structure is not identical to that of other bacterial F1FO-ATPases.The anaerobic bacteria enterococci (streptococci) have a unique proton-translocating ATPase (H+-ATPase) which does not synthesize ATP but regulates the cytoplasmic pH in this species (16,17,29). The cytoplasmic pH is regulated by proton extrusion via the H+-ATPase and by potassium influx via the specific transport system for this ion. The latter system is not absolutely necessary for pH regulation, but dissipation of the membrane potential generated by the former system is definitely required. In fact, maintenance of neutral cytoplasmic pH can be attained in the presence of valinomycin, which dissipates the membrane potential (28).Biosynthesis of H+-ATPase is stimulated at a low cytoplasmic pH (2, 31). Data from our laboratory have shown that an elevated amount of H+-ATPase at a low pH is essential for maintaining neutral cytoplasmic pH (32, 49). Thus, it now appears that the primary factor for regulating cytoplasmic pH is the H+-ATPase in enterococci (streptococci). Abrams and his coworkers (1, 3-5, 36) have shown that Enterococcus hirae (formerly Streptococcus faecalis) H+-ATPase is biochemically similar to an F1FO-ATPase of aerobic bacteria which synthesizes ATP coupled with the proton motive force.In this study, we examined the gene structure of this H+-ATPase with E. hirae ATCC 9790. The operon for E. hirae H+-ATPase contained eight genes for a, c, b, delta, alpha, gamma, beta, and epsilon subunits in that order, and the deduced amino acid sequences of E. hirae H+-ATPase subunits had a high homology with those of other bacterial F1FO-ATPases. These with 5% trichloroacetic acid for 20 min in boiling water. Trichloroacetic acid was extrac...

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We estimated the concentration of soluble IL-2R (sIL-2R) in the serum of patients with systemic lupus erythematosus (SLE) and examined the relationship between the serum levels of sIL-2R and clinical features or laboratory data. We found that elevated levels of sIL-2R were present in the serum of SLE patients with discoid rash, and sIL-2R concentrations were correlated with the soluble CD4 and soluble CD8 concentrations but not with classical serological marker, anti-DNA antibody or complement titer.

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A human B cell line, TKS-1, which was established from the peripheral blood of a patient with rheumatoid arthritis, was found to spontaneously produce a factor which enhances the activity of interleukin 1 (IL-1). This factor, designated B cell-derived growth-enhancing factor (BGEF), enhanced IL-1-induced proliferation of peanut agglutinin nonagglutinated thymocytes. BGEF also enhanced IL-1-induced production of interleukin 2 (IL-2) by both thymocytes and a human T cell clone, HSB.2 C5B2. BGEF alone did not induce the production of IL-2. BGEF failed to induce proliferation of the IL-2-dependent T cell clone, and did not enhance its response to IL-2. The activity of BGEF was not blocked by antisera against human IL-1-alpha or human IL-1-beta. Gel filtration analysis revealed that BGEF has a m.w. of 60,000 to 65,000 in its native state. We concluded that BGEF differed from IL-1 and IL-2, but is a novel factor produced by TKS-1 cells. In addition, we found that partially purified B cells from patients with rheumatoid arthritis produced factors which enhanced the activity of IL-1.