caa a22 4500 605504652 CHVBK 20210128100617.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00253-015-7012-4 doi (NATIONALLICENCE)springer-10.1007/s00253-015-7012-4 Tryptophan promotes morphological and physiological differentiation in Streptomyces coelicolor [Elektronische Daten] [Emilia Palazzotto, Giovanni Renzone, Pietro Fontana, Luigi Botta, Andrea Scaloni, Anna Puglia, Giuseppe Gallo] The molecular mechanisms regulating tryptophan biosynthesis in actinomycetes are poorly understood; similarly, the possible roles of tryptophan in the differentiation program of microorganism life-cycle are still underexplored. To unveil the possible regulatory effect of this amino acid on gene expression, an integrated study based on quantitative teverse transcription-PCR (qRT-PCR) and proteomic approaches was performed on the actinomycete model Streptomyces coelicolor. Comparative analyses on the microorganism growth in a minimal medium with or without tryptophan supplementation showed that biosynthetic trp gene expression in S. coelicolor is not subjected to a negative regulation by the presence of the end product. Conversely, tryptophan specifically induces the transcription of trp genes present in the biosynthetic gene cluster of the calcium-dependent antibiotic (CDA), a lipopeptide containing d- and l-tryptophan residues. In addition, tryptophan stimulates the transcription of the CDA gene cluster regulator cdaR and, coherently, CDA production. Surprisingly, tryptophan also promotes the production of actinorhodin, another antibiotic that does not contain this amino acid in its structure. Combined 2D-DIGE and nano liquid chromatography electrospray linear ion trap tandem mass spectrometry (LC-ESI-LIT-MS/MS) analyses revealed that tryptophan exerts a growth-stage-dependent global effect on S. coelicolor proteome, stimulating anabolic pathways and promoting the accumulation of key factors associated with morphological and physiological differentiation at the late growth stages. Phenotypic observations by scanning electron microscopy and spore production assays demonstrated an increased sporulation in the presence of tryptophan. Transcriptional analysis of catabolic genes kynA and kynB suggested that the actinomycete also uses tryptophan as a carbon and nitrogen source. In conclusion, this study originally provides the molecular basis underlying the stimulatory effect of tryptophan on the production of antibiotics and morphological development program of this actinomycete. Springer-Verlag Berlin Heidelberg, 2015 Tryptophan nationallicence S. coelicolor nationallicence Differentiation nationallicence Actinorhodin nationallicence CDA nationallicence 2D-DIGE nationallicence Palazzotto Emilia Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, 62032, Camerino, Italy aut Renzone Giovanni Proteomic and Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147, Naples, Italy aut Fontana Pietro Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128, Palermo, Italy aut Botta Luigi Dipartimento di Ingegneria Civile, Ambientale, Aerospaziale, dei Materiali, Università di Palermo, 90128, Palermo, Italy aut Scaloni Andrea Proteomic and Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147, Naples, Italy aut Puglia Anna Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128, Palermo, Italy aut Gallo Giuseppe Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128, Palermo, Italy aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 10177-10189 0175-7598 99:23<10177 2015 99 253 https://doi.org/10.1007/s00253-015-7012-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-7012-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Palazzotto Emilia Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, 62032, Camerino, Italy aut NATIONALLICENCE 700 1- Renzone Giovanni Proteomic and Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147, Naples, Italy aut NATIONALLICENCE 700 1- Fontana Pietro Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128, Palermo, Italy aut NATIONALLICENCE 700 1- Botta Luigi Dipartimento di Ingegneria Civile, Ambientale, Aerospaziale, dei Materiali, Università di Palermo, 90128, Palermo, Italy aut NATIONALLICENCE 700 1- Scaloni Andrea Proteomic and Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147, Naples, Italy aut NATIONALLICENCE 700 1- Puglia Anna Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128, Palermo, Italy aut NATIONALLICENCE 700 1- Gallo Giuseppe Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, 90128, Palermo, Italy aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 10177-10189 0175-7598 99:23<10177 2015 99 253