Dataset Information

Identification of Bacillus subtilis YjoB-binding proteins using AP-MS

ABSTRACT: Bacillus subtilis YjoB has been an uncharacterized AAA+ ATPase which consists of a single ATPase domain and a unique N-terminal domain. To understand the molecular function of YjoB, we identified YjoB-binding proteins through proteomics approaches, and analyzed the relationship between YjoB and its binding proteins. YjoB was coeluted with catalase and iron-sulfur cluster proteins in affinity purification and enhanced the activity of a catalase by specifically preventing heat-induced aggregation of catalase.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Bacillus Subtilis Subsp. Subtilis Str. 168

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Unusual Infection

SUBMITTER: Eunju Kwon

LAB HEAD: Dong Young Kim

PROVIDER: PXD032982 | Pride | 2022-10-12

REPOSITORIES: Pride

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Publications

Crystal structure and biochemical analysis suggest that YjoB ATPase is a putative substrate-specific molecular chaperone.

Kwon Eunju E Dahal Pawan P Kim Dong Young DY

Proceedings of the National Academy of Sciences of the United States of America 20221003 41

AAA+ ATPases are ubiquitous proteins associated with most cellular processes, including DNA unwinding and protein unfolding. Their functional and structural properties are typically determined by domains and motifs added to the conserved ATPases domain. Currently, the molecular function and structure of many ATPases remain elusive. Here, we report the crystal structure and biochemical analyses of YjoB, a <i>Bacillus subtilis</i> AAA+ protein. The crystal structure revealed that the YjoB hexamer ...[more]

PMID: 36191235

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Project description:Cilia assembly, maintenance and the localization of signal-transduction proteins necessitate a functioning intraflagellar transport (IFT). The IFT machinery moves cargo along a microtubule scaffold from the proximal to the distal end of the cilium. This anterograde transport is facilitated by a large multiprotein complex IFT-B, which consists of 16 proteins in total and recruits motor protein Kinesin-2 for active movement. TTC30A and TTC30B are integral components of this complex and were shown before to have redundant function in context of IFT. One paralogue could compensate the loss of the other, preventing the disruption of IFT-B and thus a severe ciliogenesis defect with no formation of the cilium. Affinity-based protein complex analysis of endogenously tagged cells, generated via CRISPR/Cas9 system as described before, revealed paralogue specific protein interactors proposing the involvement of TTC30A or TTC30B in other ciliary functions, particularly Sonic hedgehog signaling (Shh). Defects in this ciliary signaling pathway are often correlated to synpolydactyly, which intriguingly is also linked to a rare TTC30 variant. In this study we focus on the so far unknown interaction of TTC30A with protein kinase A catalytic subunit α PRKACA, which is a negative regulator of Shh pathway. For an in-depth analysis of this unique interaction and the influence on Shh, we used previously, via the CRISPR/Cas9 system, generated TTC30A or B single- and double-knockout hTERT-RPE1 cells as well as rescue cells harboring the TTC30 mutation. Our systematic approach revealed the paralogue specific influence of TTC30A KO and mutated TTC30A on the activity of PRKACA as well as the uptake of Smoothened into the cilium resulting in a downregulation of Sonic hedgehog signaling. Affinity-based protein complex analysis of wildtype versus mutated TTC30A or TTC30B uncovered differences in interaction pattern. These interactome alterations combined with Shh downregulation suggest a possible mechanism of how mutant TTC30A and respectively TTC30A KO are linked to synpolydactyly.

Dataset Information

Identification of Bacillus subtilis YjoB-binding proteins using AP-MS

Publications

Crystal structure and biochemical analysis suggest that YjoB ATPase is a putative substrate-specific molecular chaperone.

Similar Datasets

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