PUL ID

PUL0195

PubMed

26691835, Mol Microbiol. 2016 Apr;100(2):289-302. doi: 10.1111/mmi.13316. Epub 2016 Feb 9.

Characterization method

RT-PCR

Genomic accession number

NC_003030.1

Nucelotide position range

444719-451147

Substrate

cellobiose

Loci

CA_C0382-CA_C0386; CelRCA-bglB-celC

Species

Clostridium acetobutylicum/1488

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

levR CA_RS02160 WP_010963704.1 0 - 2691 (-) NC_003030.1:444719-447410 -
- CA_RS02165 WP_010963705.1 2931 - 3252 (+) NC_003030.1:447650-447971 -
licB CA_RS02170 WP_010963706.1 3277 - 3586 (+) NC_003030.1:447996-448305 -
- CA_RS02175 WP_010963707.1 3610 - 5020 (+) NC_003030.1:448329-449739 -
licC CA_RS02180 WP_010963708.1 5076 - 6429 (+) NC_003030.1:449795-451148 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

levR 1 - 2691 (-) TC: gnl|TC-DB|Q9KN48|3.A.23.1.1 Yes
- 2932 - 3252 (+) TC: gnl|TC-DB|Q045H4|4.A.3.1.3 Yes
licB 3278 - 3586 (+) TC: gnl|TC-DB|Q72XP9|4.A.3.2.8 Yes
- 3611 - 5020 (+) CAZyme: GH1 Yes
licC 5077 - 6429 (+) TC: gnl|TC-DB|Q72XQ0|4.A.3.2.8 Yes

PUL ID

PUL0195

PubMed

26691835, Mol Microbiol. 2016 Apr;100(2):289-302. doi: 10.1111/mmi.13316. Epub 2016 Feb 9.

Title

PTS regulation domain-containing transcriptional activator CelR and sigma factor sigma(54) control cellobiose utilization in Clostridium acetobutylicum.

Author

Nie X, Yang B, Zhang L, Gu Y, Yang S, Jiang W, Yang C

Abstract

The phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) regulation domain (PRD)-containing enhancer binding proteins (EBPs) are an important class of sigma(54) -interacting transcriptional activators. Although PRD-containing EBPs are present in many Firmicutes, most of their regulatory functions remain unclear. In this study, the transcriptional regulons of about 50 PRD-containing EBPs in diverse Firmicutes species are reconstructed by using a comparative genomic approach, which contain the genes associated with utilization of beta-glucosides, fructose/levan, mannose/glucose, pentitols, and glucosamine/fructosamine. We then present experimental evidence that the cel operon involved in cellobiose utilization is directly regulated by CelR and sigma(54) (SigL) in Clostridium acetobutylicum. The predicted three CelR-binding sites and sigma(54) promoter elements upstream of the cel operon are verified by in vitro binding assays. We show that CelR has an ATPase activity, which is strongly stimulated by the presence of DNA containing the CelR-binding sites. Moreover, mutations in any one of the three CelR-binding sites significantly decreased the cel promoter activity probably due to the need for all three DNA sites for maximal ATPase activity of CelR. It is suggested that CelR is regulated by PTS-mediated phosphorylation at His-551 and His-829, which exerts a positive effect and an inhibitory effect, respectively, on the CelR activity.