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Volume 13
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10.3390/genes13071191
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Article

TTC30A and TTC30B Redundancy Protects IFT Complex B Integrity and Its Pivotal Role in Ciliogenesis

by
Felix Hoffmann
1,
Sylvia Bolz
1,
Katrin Junger
1,
Franziska Klose
1,
Timm Schubert
1,2,
Franziska Woerz
1,
Karsten Boldt
1,
Marius Ueffing
1 and
Tina Beyer
1,*
1
Institute for Ophthalmic Research, Eberhard Karls University Tübingen, Elfriede-Aulhorn-Str. 7, 72076 Tübingen, Germany
2
Werner Reichardt Centre for Integrative Neuroscience (CIN), University of Tübingen, Otfried-Müller-Str. 25, 72076 Tübingen, Germany
*
Author to whom correspondence should be addressed.
Genes 2022, 13(7), 1191; https://doi.org/10.3390/genes13071191
Submission received: 29 April 2022 / Revised: 7 June 2022 / Accepted: 29 June 2022 / Published: 1 July 2022
(This article belongs to the Special Issue Updates on Cilia Biology and Cilia-Associated Disorders)
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Abstract

Intraflagellar transport (IFT) is a microtubule-based system that supports the assembly and maintenance of cilia. The dysfunction of IFT leads to ciliopathies of variable severity. Two of the IFT-B components are the paralogue proteins TTC30A and TTC30B. To investigate whether these proteins constitute redundant functions, CRISPR/Cas9 was used to generate single TTC30A or B and double-knockout hTERT-RPE1 cells. Ciliogenesis assays showed the redundancy of both proteins while the polyglutamylation of cilia was affected in single knockouts. The localization of other IFT components was not affected by the depletion of a single paralogue. A loss of both proteins led to a severe ciliogenesis defect, resulting in no cilia formation, which was rescued by TTC30A or B. The redundancy can be explained by the highly similar interaction patterns of the paralogues; both equally interact with the IFT-B machinery. Our study demonstrates that a loss of one TTC30 paralogue can mostly be compensated by the other, thus preventing severe ciliary defects. However, cells assemble shorter cilia, which are potentially limited in their function, especially because of impaired polyglutamylation. A complete loss of both proteins leads to a deficit in IFT complex B integrity followed by disrupted IFT and subsequently no cilia formation.
Keywords: cilia; IFT; TTC30 paralogues; CRISPR/Cas9; affinity proteomics; polyglutamylation cilia; IFT; TTC30 paralogues; CRISPR/Cas9; affinity proteomics; polyglutamylation
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MDPI and ACS Style

Hoffmann, F.; Bolz, S.; Junger, K.; Klose, F.; Schubert, T.; Woerz, F.; Boldt, K.; Ueffing, M.; Beyer, T. TTC30A and TTC30B Redundancy Protects IFT Complex B Integrity and Its Pivotal Role in Ciliogenesis. Genes 2022, 13, 1191. https://doi.org/10.3390/genes13071191

AMA Style

Hoffmann F, Bolz S, Junger K, Klose F, Schubert T, Woerz F, Boldt K, Ueffing M, Beyer T. TTC30A and TTC30B Redundancy Protects IFT Complex B Integrity and Its Pivotal Role in Ciliogenesis. Genes. 2022; 13(7):1191. https://doi.org/10.3390/genes13071191

Chicago/Turabian Style

Hoffmann, Felix, Sylvia Bolz, Katrin Junger, Franziska Klose, Timm Schubert, Franziska Woerz, Karsten Boldt, Marius Ueffing, and Tina Beyer. 2022. "TTC30A and TTC30B Redundancy Protects IFT Complex B Integrity and Its Pivotal Role in Ciliogenesis" Genes 13, no. 7: 1191. https://doi.org/10.3390/genes13071191

APA Style

Hoffmann, F., Bolz, S., Junger, K., Klose, F., Schubert, T., Woerz, F., Boldt, K., Ueffing, M., & Beyer, T. (2022). TTC30A and TTC30B Redundancy Protects IFT Complex B Integrity and Its Pivotal Role in Ciliogenesis. Genes, 13(7), 1191. https://doi.org/10.3390/genes13071191

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