Molecular and functional analyses of the gene (eshA) encoding the 52-kilodalton protein of Streptomyces coelicolor A3(2) required for antibiotic production.

Analysis of proteins recovered in the S100 precipitate fraction of Streptomyces griseus after ultracentrifugation led to the identification of a 52-kDa protein which is produced during the late growth phase. The gene (eshA) which codes for this protein was cloned from S. griseus, and then its homologue was cloned from Streptomyces coelicolor A3(2). The protein was deduced to be 471 amino acids in length. The protein EshA is characterized by a central region that shows homology to the eukaryotic-type cyclic nucleotide-binding domains. Significant homology was also found to MMPI in Mycobacterium leprae, a major antigenic protein to humans. The eshA gene mapped near the chromosome end and was not essential for viability, as demonstrated by gene disruption experiments, but its disruption resulted in the abolishment of an antibiotic (actinorhodin but not undecylprodigiosin) production. Aerial mycelium was produced as abundantly as by the parent strain. Expression analysis of the EshA protein by Western blotting revealed that EshA is present only in late-growth-phase cells. The eshA gene was transcribed just preceding intracellular accumulation of the EshA protein, as determined by S1 nuclease protection, indicating that EshA expression is regulated at the transcription level. The expression of EshA was unaffected by introduction of the relA mutation, which blocks ppGpp synthesis.

[Epidemiological immunological studies on leprosy in Okinawa. 6. Lepromin, lymphocyte transformation, FLA-ABS and leproagglutination tests in the in-and out-patients with leprosy and the relationships among these tests and personal or family history or clinical findings of the patients].

In- and out-patients with leprosy in Okinawa were surveyed by using the following immunological tests: cutaneous reaction to Mitsuda's lepromin (40 million bacilli/ml) (LPR), lymphocyte transformation test with Dharmendara's antigen (DL-LTT), that with phytohemagglutinin (PHA-LTT), fluorescent leprosy antibody absorption test (FLA-ABS) and Leproagglutination test with cardiolipin-lecithin antigen (LAT). A correlation between two tests and a relationship between the test and personal or family history or clinical finding of the patients were evaluated by non-parametric statistics. Spearman's rank correlation coefficient was significant between LPR and DL-LTT, while a reverse correlation was found between LPR and FLA-ABS, DL-LTT and FLA-ABS, and DL-LTT and LAT, respectively. High reactivity in PHA-LTT was found in almost all of patients, irrespective of type and stage of leprosy, suggesting general immune responsiveness of leprosy patients in Okinawa. The other tests showed a significant difference in the mean rank of reaction values according to clinical findings such as type and stage of leprosy, property of skin lesion, loss of eyebrow, and bacteriological test. The use of LPR and DL-LTT for the study of cell-mediated immunity in leprosy was therefore confirmed. FLA-ABS was found useful for early serodiagnosis, while LAT for the study of autoimmunity in leprosy. None of these immunological tests showed significant difference among the subgroups of patients classified by the following items: presence or absence of consanguineous patient with leprosy, living place, place of onset, presence or absence of neural symptoms such as the enlargement of peripheral nerve, claw hand and facial palsy, ocular and nasal involvement, ENL in lepromatous and borderline leprosy and conditions in the treatment. On the other hand, LPR showed higher reactivity in female than in male, in the patients at home than those discharged, and in the patients with drop foot than those without. Higher reactivity of DL-LTT was also found in male than in female and in the patients with plantar ulcer than those without. FLA-ABS reactivity was significantly higher in the discharged patients than the inpatients, in the period of 5 to 9 years after onset than that of 20 or more years, but the reactivity was lower in the patients with drop foot than those without. LAT reactivity was significantly higher in the 30-39 years old patients than 0-19 years old, and in the patients with physical work than those with mental.(ABSTRACT TRUNCATED AT 400 WORDS)