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- Title
The Signaling Pathway That cGAMP Riboswitches Found: Analysis and Application of Riboswitches to Study cGAMP Signaling in Geobacter sulfurreducens.
- Authors
Tan, Zhesen; Chan, Chi Ho; Maleska, Michael; Banuelos Jara, Bryan; Lohman, Brian K.; Ricks, Nathan J.; Bond, Daniel R.; Hammond, Ming C.
- Abstract
The Hypr cGAMP signaling pathway was discovered via the function of the riboswitch. In this study, we show the development of a method for affinity capture followed by sequencing to identify non-coding RNA regions that bind nucleotide signals such as cGAMP. The RNAseq of affinity-captured cGAMP riboswitches from the Geobacter sulfurreducens transcriptome highlights general challenges that remain for this technique. Furthermore, by applying riboswitch reporters in vivo, we identify new growth conditions and transposon mutations that affect cGAMP levels in G. sulfurreducens. This work reveals an extensive regulatory network and supports a second functional cGAMP synthase gene in G. sulfurreducens. The activity of the second synthase was validated using riboswitch-based fluorescent biosensors, and is the first known example of an active enzyme with a variant GGDDF motif.
- Subjects
GEOBACTER sulfurreducens; RIBOSWITCHES; CELLULAR signal transduction; NON-coding RNA; SOCIAL networks
- Publication
International Journal of Molecular Sciences, 2022, Vol 23, Issue 3, p1183
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms23031183