TrpM, a Small Protein Modulating Tryptophan Biosynthesis and Morpho-Physiological Differentiation in Streptomyces coelicolor A3(2).
In the model actinomycete Streptomyces coelicolor A3(2), small open reading frames encoding proteins with unknown functions were identified in several amino acid biosynthetic gene operons, such as SCO2038 (trpX) in the tryptophan trpCXBA locus. In this study, the role of the corresponding protein in tryptophan biosynthesis was investigated by combining phenotypic and molecular analyses. The 2038KO mutant strain was characterized by delayed growth, smaller aerial hyphae and reduced production of spores and actinorhodin antibiotic, with respect to the WT strain. The capability of this mutant to grow on minimal medium was rescued by tryptophan and tryptophan precursor (serine and/or indole) supplementation on minimal medium and by gene complementation, revealing the essential role of this protein, here named TrpM, as modulator of tryptophan biosynthesis. His-tag pull-down and bacterial adenylate cyclase-based two hybrid assays revealed TrpM interaction with a putative leucyl-aminopeptidase (PepA), highly conserved component among various Streptomyces spp. In silico analyses showed that PepA is involved in the metabolism of serine, glycine and cysteine through a network including GlyA, CysK and CysM enzymes. Proteomic experiments suggested a TrpM-dependent regulation of metabolic pathways and cellular processes that includes enzymes such as GlyA, which is required for the biosynthesis of tryptophan precursors and key proteins participating in the morpho-physiological differentiation program. Altogether, these findings reveal that TrpM controls tryptophan biosynthesis at the level of direct precursor availability and, therefore, it is able to exert a crucial effect on the morpho-physiological differentiation program in S. coelicolor A3(2).
Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, 62032 Camerino, Italy.|Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128 Palermo, Italy.
Emilia
E
Palazzotto
Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128 Palermo, Italy.
Giuseppe
G
Gallo
Proteomic and Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147 Naples, Italy.
Giovanni
G
Renzone
Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128 Palermo, Italy.
Anna
A
Giardina
Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128 Palermo, Italy.
Alberto
A
Sutera
Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128 Palermo, Italy.
Joohee
J
Silva
Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128 Palermo, Italy.
Celinè
C
Vocat
Dipartimento di Ingegneria Civile, Ambientale, Aerospaziale, dei Materiali, University of Palermo, 90128 Palermo, Italy.
Luigi
L
Botta
Proteomic and Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147 Naples, Italy.
Andrea
A
Scaloni
Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128 Palermo, Italy.
Anna Maria
AM
Puglia
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