Universität zu Köln

Uhde, Andreas, Bruehl, Natalie, Goldbeck, Oliver ORCID: 0000-0002-5708-8600, Matano, Christian, Gurow, Oksana, Rueckert, Christian, Marin, Kay, Wendisch, Volker F. ORCID: 0000-0003-3473-0012, Kraemer, Reinhard and Seibold, Gerd M. (2016). Transcription of Sialic Acid Catabolism Genes in Corynebacterium glutamicum Is Subject to Catabolite Repression and Control by the Transcriptional Repressor NanR. J. Bacteriol., 198 (16). S. 2204 - 2219. WASHINGTON: AMER SOC MICROBIOLOGY. ISSN 1098-5530

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Abstract

Corynebacterium glutamicum metabolizes sialic acid (Neu5Ac) to fructose-6-phosphate (fructose-6P) via the consecutive activity of the sialic acid importer SiaEFGI, N-acetylneuraminic acid lyase (NanA), N-acetylmannosamine kinase (NanK), N-acetylmannosamine- 6P epimerase (NanE), N-acetylglucosamine-6P deacetylase (NagA), and glucosamine-6P deaminase (NagB). Within the cluster of the three operons nagAB, nanAKE, and siaEFGI for Neu5Ac utilization a fourth operon is present, which comprises cg2936, encoding a GntR-type transcriptional regulator, here named NanR. Microarray studies and reporter gene assays showed that nagAB, nanAKE, siaEFGI, and nanR are repressed in wild-type (WT) C. glutamicum but highly induced in a Delta nanR C. glutamicum mutant. Purified NanR was found to specifically bind to the nucleotide motifs A[AC] G[CT][AC] TGATGT C[AT][TG] ATGT[AC] TA located within the nagA-nanA and nanR-sialA intergenic regions. Binding of NanR to promoter regions was abolished in the presence of the Neu5Ac metabolism intermediates GlcNAc-6P and N-acetylmannosamine-6-phosphate (ManNAc-6P). We observed consecutive utilization of glucose and Neu5Ac as well as fructose and Neu5Ac by WT C. glutamicum, whereas the deletion mutant C. glutamicum Delta nanR simultaneously consumed these sugars. Increased reporter gene activities for nagAB, nanAKE, and nanR were observed in cultivations of WT C. glutamicum with Neu5Ac as the sole substrate compared to cultivations when fructose was present. Taken together, our findings show that Neu5Ac metabolism in C. glutamicum is subject to catabolite repression, which involves control by the repressor NanR. IMPORTANCE Neu5Ac utilization is currently regarded as a common trait of both pathogenic and commensal bacteria. Interestingly, the nonpathogenic soil bacterium C. glutamicum efficiently utilizes Neu5Ac as a substrate for growth. Expression of genes for Neu5Ac utilization in C. glutamicum is here shown to depend on the transcriptional regulator NanR, which is the first GntR-type regulator of Neu5Ac metabolism not to use Neu5Ac as effector but relies instead on the inducers GlcNAc-6P and ManNAc-6P. The identification of conserved NanR-binding sites in intergenic regions within the operons for Neu5Ac utilization in pathogenic Corynebacterium species indicates that the mechanism for the control of Neu5Ac catabolism in C. glutamicum by NanR as described in this work is probably conserved within this genus.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Uhde, Andreas UNSPECIFIED UNSPECIFIED UNSPECIFIED Bruehl, Natalie UNSPECIFIED UNSPECIFIED UNSPECIFIED Goldbeck, Oliver UNSPECIFIED orcid.org/0000-0002-5708-8600 UNSPECIFIED Matano, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Gurow, Oksana UNSPECIFIED UNSPECIFIED UNSPECIFIED Rueckert, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Marin, Kay UNSPECIFIED UNSPECIFIED UNSPECIFIED Wendisch, Volker F. UNSPECIFIED orcid.org/0000-0003-3473-0012 UNSPECIFIED Kraemer, Reinhard UNSPECIFIED UNSPECIFIED UNSPECIFIED Seibold, Gerd M. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-268881
DOI:	10.1128/JB.00820-15
Journal or Publication Title:	J. Bacteriol.
Volume:	198
Number:	16
Page Range:	S. 2204 - 2219
Date:	2016
Publisher:	AMER SOC MICROBIOLOGY
Place of Publication:	WASHINGTON
ISSN:	1098-5530
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DNA-BINDING SITES; ESCHERICHIA-COLI; STREPTOCOCCUS-PNEUMONIAE; N-ACETYLMANNOSAMINE; RECEPTOR PROTEIN; GENOME SEQUENCE; GLUCOSE-UPTAKE; AMINO-ACIDS; EXPRESSION; GROWTH Multiple languages Microbiology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/26888