Abstract
Corynebacterium glutamicum WhcD plays an important regulatory role in cell division. Binding of WhcD to the promoter region of its target genes, such as ftsZ, was observed by electrophoretic mobility shift assays (EMSA) using purified fusion proteins; however, binding could only be observed in the presence of WhiA. Although WhcD alone did not bind to the DNA, it stimulated binding of WhiA to the promoter region of the cell division gene ftsZ. Binding of WhcD and WhiA to DNA did not occur in the presence of the oxidant diamide. Purified WhcD and WhiA physically interacted in vitro. The presence of diamide did not disrupt the WhcD–WhiA interaction but affected binding of WhiA to the promoter region of ftsZ. The GACAC motif and adjacent sequences were found to be important for binding of the WhcD–WhiA complex to the DNA. Collectively, our results suggest that WhcD enhances the WhiA DNA-binding activity by physically interacting with WhiA. In addition, loss of WhiA DNA-binding activity in the presence of an oxidant agent may suggest a role for this protein as a switch that controls cell division in cells under oxidative stress.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Aínsa JA, Ryding NJ, Hartley N, Findlay KC, Bruton CJ, Chater KF (2000) WhiA, a protein of unknown function conserved among gram-positive bacteria, is essential for sporulation in Streptomyces coelicolor A3(2). J Bacteriol 182:5470–5478
Bush MJ, Bibb MJ, Chandra G, Findlay KC, Buttner MJ (2013) Genes required for aerial growth, cell division, and chromosome segregation are targets of WhiA before sporulation in Streptomyces venezuelae. MBio 4:e00684–e00713
Bush MJ, Chandra G, Bibb MJ, Findlay KC, Buttner MJ (2016) Genome-wide chromatin immunoprecipitation sequencing analysis shows that WhiB is a transcription factor that cocontrols its regulon with WhiA to initiate developmental cell division in Streptomyces. MBio 7:e00523–e00616
Chater KF (1975) Construction and phenotypes of double sporulation deficient mutants in Streptomyces coelicolor A3(2). J Gen Microbiol 87:312–325
Chater KF, Chandra G (2006) The evolution of development in Streptomyces analysed by genome comparisons. FEMS Microbiol Rev 30:651–672
Choi WW, Park SD, Lee SM, Kim HB, KimY Lee HS (2009) The whcA gene plays a negative role in oxidative stress response of Corynebacterium glutamicum. FEMS Microbiol Lett 290:32–38
Crack JC, Smith LJ, Stapleton MR, Peck J, Watmough NJ, Buttner MJ, Buxton RS, Green J, Oganesyan VS, Thomson AJ (2010) Mechanistic insight into the nitrosylation of the [4Fe-4S] cluster of WhiB-like proteins. J Am Chem Soc 133:1112–1121
Crack JC, Munnoch J, Dodd EL, Knowles F, Al Bassam MM, Kamali S, Holland AA, Cramer SP, Hamilton CJ, Johnson MK, Thomson AJ, Hutchings MI, Le Brun NE (2015) NsrR from Streptomyces coelicolor is a nitric oxide-sensing [4Fe-4S] cluster protein with a specialized regulatory function. J Biol Chem 290:12689–12704
den Hengst CD, Buttner MJ (2008) Redox control in Actinobacteria. Biochimica et Biophysica Acta (BBA) 1780:1201–1216
Flärdh K, Findlay KC, Chater KF (1999) Association of early sporulation genes with suggested developmental decision points in Streptomyces coelicolor A3(2). Microbiology 145:2229–2243
Follettie MT, Sinskey AJ (1986) Recombinant DNA technology for Corynebacterium glutamicum. Food Technol 40:88–94
Grant SG, Jessee J, Bloom FR, Hanahan D (1990) Differential plasmid rescue from transgenic mouse DNAs into Escherichia coli methylation-restriction mutants. Proc Natl Acad Sci USA 87:4645–4649
Kaiser BK, Stoddard BL (2011) DNA recognition and transcriptional regulation by the WhiA sporulation factor. Sci Rep 1:156
Kaiser BK, Clifton MC, Shen BW, Stoddard BL (2009) The structure of a bacterial DUF199/WhiA protein: domestication of an invasive endonuclease. Structure 17:1368–1376
Kim TH, Park JS, Kim HJ, Kim Y, Kim P, Lee HS (2005) The whcE gene of Corynebacterium glutamicum is important for survival following heat and oxidative stress. Biochem Biophys Res Commun 337:757–764
Knizewski L, Ginalski K (2007) Bacterial DUF199/COG1481 proteins including sporulation regulator WhiA are distant homologs of LAGLIDADG homing endonucleases that retained only DNA binding. Cell Cycle 6:1666–1670
Lee JY, Park JS, Kim HJ, Kim Y, Lee HS (2012) Corynebacterium glutamicum whcB, a stationary phase-specific regulatory gene. FEMS Microbiol Lett 327:103–109
Lee JY, Na YA, Kim E, Lee HS, Kim P (2016) The Actinobacterium Corynebacterium glutamicum, an industrial workhorse. J Microbiol Biotechnol 26:807–822
Lee DS, Kim Y, Lee HS (2017) The whcD gene of Corynebacterium glutamicum plays roles in cell division and envelope formation. Microbiology 163:131–143
Letek M, Fiuza M, Ordóñez E, Villadangos AF, Ramos A, Mateos LM, Gil JA (2008) Cell growth and cell division in rod-shaped actinomycete Corynebacterium glutamicum. Antonie Van Leeuwenhoek 94:99–109
Park SD, Youn JW, Kim YJ, Lee SM, Kim Y, Lee HS (2008) Corynebacterium glutamicum σ E is involved in responses to cell surface stresses and its activity is controlled by the anti-σ factor CseE. Microbiology 154:915–923
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Schwedock J, McCormick JR, Angert ER, Nodwell JR, Losick R (1997) Assembly of the cell division protein FtsZ into ladder-like structures in the aerial hyphae of Streptomyces coelicolor. Mol Microbiol 25:847–858
Singh A, Crossman DK, Mai D, Guidry L, Voskuil MI, Renfrow MB, Steyn AJC (2009) Mycobacterium tuberculosis WhiB3 maintains redox homeostasis by regulating virulence lipid anabolism to modulate macrophage response. PLoS Pathog 5:e1000545
Yoshihama M, Higashiro K, Rao EA, Akedo M, Shanabruch WG, Follettie MT, Walker GC, Sinskey AJ (1985) Cloning vector system for Corynebacterium glutamicum. J Bacteriol 162:591–597
Zheng F, Long Q, Xie J (2012) The function and regulatory network of WhiB and WhiB-like protein from comparative genomics and systems biology perspectives. Cell Biochem Biophys 63:103–108
Acknowledgements
This work was supported by the Basic Science Research Program, through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2016R1D1A3A03916638), and in part by the Framework of International Cooperation Program, through the National Research Foundation of Korea (NRF) funded by the Ministry of Science (NRF-2016K2A9A2A10005545).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Lee, DS., Kim, P., Kim, ES. et al. Corynebacterium glutamicum WhcD interacts with WhiA to exert a regulatory effect on cell division genes. Antonie van Leeuwenhoek 111, 641–648 (2018). https://doi.org/10.1007/s10482-017-0953-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10482-017-0953-0