Genetic analysis of sulfate assimilation gene cluster of Streptomyces coelicolor A3(2)

  • B. O. Ostash Ivan Franko National University of Lviv, Ukraine, 79005, Lviv, Hrushevskoho, 4 https://orcid.org/0000-0001-5904-5957
  • K. Stanchak Ivan Franko National University of Lviv, Ukraine, 79005, Lviv, Hrushevskoho, 4
  • T. Gren Ivan Franko National University of Lviv, Ukraine, 79005, Lviv, Hrushevskoho, 4; The Novo Nordisk Foundation Center for Biosustainability of DTU, Denmark, 2800, Kgs. Lyngby
  • V. O. Fedorenko Ivan Franko National University of Lviv, Ukraine, 79005, Lviv, Hrushevskoho, 4
Keywords: Streptomyces coelicolor A3(2), genetics of sulfur metabolism, sulfate assimilation

Abstract

Aim. Streptomyces coelicolor A3(2) is the best studied species within this bacterial genus. Biosynthesis of specialized (secondary) metabolites by Streptomyces is of special interest. Primary metabolism, where all the precursors of specialized metabolites come from, is also studied in great detail. There are glaring gaps in our knowledge of sulfur metabolism in this species. We took genetic approach to probe the function of several genes within presumed sulfate assimilation gene cluster of S. coelicolor A3(2). Methods. Microbiological and genetic approaches were combined to generate mutants and to study their properties. Results. Sulfate assimilation gene cluster is structurally and functionally similar to that of phylogenetically close Corynebacterium. Most of the generated knockout strains behaved as would be expected from their molecular function inferred in silico. This confirms their involvement in sulfate uptake/conversion. Knockout of gene sco6101 (having no homologs from the other bacterial sulfate assimilation operons) impaired the growth on inorganic sulfur species and L-cysteine, pointing to its association with sulfur metabolism. Conclusions. Our study provides experimental evidence for the involvement of sco6093-sco6102 segment in sulfate assimilation, and also reveals novel gene, sco6101, essential for sulfur cycle. Further efforts are needed to elucidate the mechanism of Sco6101 action.

References

Hodgson D. Primary metabolism and its control in streptomycetes: a most unusual group of bacteria. Adv. Microb. Physiol. 2000. Vol. 42. P. 47–238. doi: 10.1016/s0065-2911(00)42003-5.

Rückert C., Koch D., Rey D., Albersmeier A., Mormann S., Pühler A., Kalinowski J. Functional genomics and expression analysis of the Corynebacterium glutamicum fpr2-cysIXHDNYZ gene cluster involved in assimilatory sulphate reduction. BMC Genomics. 2005. Vol. 6. P. 121. doi: 10.1186/1471-2164-6-121.

Chang Z., Vining L. Biosynthesis of sulfur-containing amino acids in Streptomyces venezuelae ISP5230: roles for cystathionine beta-synthase and transsulfuration. Microbiology. 2002. Vol. 148. P. 2135–2147. doi: 10.1099/00221287-148-7-2135.

Gren T., Ostash B., Babiy V., Rokytskyy I., Fedorenko V. Analysis of Streptomyces coelicolor M145 genes SCO4164 and SCO5854 encoding putative rhodaneses. Folia Microbiol. 2018. Vol. 63. P. 197–201. doi: 10.1007/s12223-017-0551-6.

Lydiate D., Mendez C., Kieser H., Hopwood D. Mutation and cloning of clustered Streptomyces genes essential for sulfate metabolism. Mol. Gen. Genet. 1988. Vol. 211. P. 415–423. doi: 10.1007/BF00425694.

Lu T., Wu X., Cao Q., Xia Y., Xun L., Liu H. Sulfane sulfur posttranslationally modifies the global regulator AdpA to influence actinorhodin production and morphological differentiation of Streptomyces coelicolor. mBio. 2022. Vol. 13. P. e0386221. doi: 10.1128/mbio.03862-21.

Gust B., Chandra G., Jakimowicz D., Yuqing T., Bruton C., Chater K. Lambda red-mediated genetic manipulation of antibiotic-producing Streptomyces. Adv. Appl. Microbiol. 2004. Vol. 54. P. 107–128. doi: 10.1016/S0065-2164(04)54004-2.

Siegl T., Luzhetskyy A. Actinomycetes genome engineering approaches. Antonie Van Leeuwenhoek. 2012. Vol. 102. P. 503–516.

Fischer M., Schmidt C., Falke D., Sawers R. Terminal reduction reactions of nitrate and sulfate assimilation in Streptomyces coelicolor A3(2): identification of genes encoding nitrite and sulfite reductases. Res. Microbiol. 2012. Vol. 163. P. 340–348. doi: 10.1016/j.resmic.2012.05.004.

Ouchi T., Tomita T., Horie A., Yoshida A., Takahashi K., Nishida H., Lassak K., Taka H., Mineki R., Fujimura T., Kosono S., Nishiyama C., Masui R., Kuramitsu S., Albers V., Kuzuyama T., Nishiyama M. Lysine and arginine biosyntheses mediated by a common carrier protein in Sulfolobus. Nat. Chem. Biol. 2013. Vol. 9. P. 277–283. doi: 10.1038/nchembio.1200.