PXD014421 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | A TetR-family protein activates transcription from a new promoter motif associated with essential genes for autotrophic growth in acetogens |
Description | Acetogens can fix carbon (CO or CO2) into acetyl-CoA via the Wood-Ljungdahl pathway (WLP) that also makes them attractive cell factories for the production of fuels and chemicals from waste feedstocks. Although most biochemical details of the WLP are well understood and systems-level characterisation of acetogen metabolism has recently improved, key transcriptional features such as promoter motifs and transcriptional regulators are still unknown in acetogens. Here, we use differential RNA-sequencing to identify a previously undescribed promoter motif associated with essential genes for autotrophic growth of the model-acetogen Clostridium autoethanogenum. RNA polymerase was shown to bind to the new promoter motif using a DNA-binding protein assay and proteomics enabled the discovery of four candidates to potentially function directly in control of transcription of the WLP and other key genes of C1 fixation metabolism. Next, in vivo experiments showed that a TetR-family transcriptional regulator (CAETHG_0459) and the housekeeping sigma factor (σA) activate expression of a reporter protein (GFP) in-frame with the new promoter motif from a fusion vector in E. coli. Lastly, a protein-protein interaction assay with the RNA polymerase (RNAP) shows that CAETHG_0459 directly binds to the RNAP. Together, the data presented here advance the fundamental understanding of transcriptional regulation of C1 fixation in acetogens and provide a strategy for improving the performance of gas-fermenting bacteria by genetic engineering. |
HostingRepository | PRIDE |
AnnounceDate | 2019-11-20 |
AnnouncementXML | Submission_2019-11-20_06:45:45.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Kaspar Valgepea |
SpeciesList | scientific name: Clostridium autoethanogenum; NCBI TaxID: 84023; |
ModificationList | iodoacetamide derivatized residue |
Instrument | TripleTOF 5600; LTQ Orbitrap Elite |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2019-06-28 02:05:58 | ID requested | |
⏵ 1 | 2019-11-20 06:45:46 | announced | |
2 | 2019-11-27 03:03:00 | announced | 2019-11-27: Updated project metadata. |
3 | 2020-01-14 07:03:14 | announced | 2019-11-27: Updated project metadata.
2020-01-14: Updated publication reference for PubMed record(s): 31803150. |
Publication List
Dataset with its publication pending |
Keyword List
submitter keyword: biological |
data-dependent acquisition |
gas fermentation |
acetogen |
Contact List
Esteban Marcellin |
contact affiliation | Australian Institute for Bioengineering and Nanotechnolog (AIBN), The University of Queensland |
contact email | e.marcellin@uq.edu.au |
lab head | |
Kaspar Valgepea |
contact affiliation | Australian Institute for Bioengineering and Nanotechnology,The University of Queensland |
contact email | k.valgepea@uq.edu.au |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD014421
- Label: PRIDE project
- Name: A TetR-family protein activates transcription from a new promoter motif associated with essential genes for autotrophic growth in acetogens