A rare leucine codon in adpA is implicated in the morphological defect of bldA mutants of Streptomyces coelicolor

Streptomycetes are mycelial bacteria that produce sporulating aerial hyphae on solid media. Bald (bld) mutants fail to form aerial mycelium under at least some conditions. bldA encodes the only tRNA species able to read the leucine codon UUA efficiently, implying the involvement of a TTA-containing gene in initiating aerial growth. One candidate for such a gene was bldH, because the bldH109 mutant of Streptomyces coelicolor resembles bldA mutants in some aspects. In the work reported here, adpA(c), an S. coelicolor gene similar to the Streptomyces griseus A factor-regulated adpA(g), was found to complement the bldH109 mutant partially at both single and multiple copies. The sequence of adpA(c) from the bldH109 mutant revealed a frameshift. A constructed in frame deletion of adpA(c) conferred a bald colony phenotype, and the mutant behaved like bldA mutants and bldH109 in its pattern of extracellular signal exchange. Both adpA(c) and adpA(g) contain a TTA codon. A TTA-free version of adpA(c) was engineered by replacing the TTA leucine codon with a cognate TTG leucine codon. The adpA(TTA-->TTG) gene could partially restore aerial mycelium formation to a bldA mutant when it was followed in cis by the gene ornA, as in the natural chromosomal arrangement. This indicated that the UUA codon in adpA(c) mRNA is the principal target through which bldA influences morphological differentiation. It also implied that translational arrest at the UUA codon in adpA(c) mRNA caused a polar effect on the downstream ornA, and that the poor translation of both genes contributes extensively to the deficiency of aerial mycelium formation in bldA mutants. Unlike the situation in S. griseus, adpA(c) transcription does not depend on the host's gamma-butyrolactone signalling system, at least in liquid cultures. In addition, sigma factor BldN, which is the homologue of an S. griseus sigma factor AdsA that is absent from adpA(g) mutants of S. griseus, was present in the constructed adpA(c) null mutant of S. coelicolor.