Abstract
The intraflagellar transport (IFT) machinery plays a crucial role in the bidirectional trafficking of components necessary for ciliary signaling, such as the Hedgehog, Wnt/PCR, and cAMP/PKA systems. Defects in some components of the IFT machinery cause dysfunction, leading to a wide range of human diseases and developmental disorders termed ciliopathies, such as nephronophthisis. The IFT machinery comprises three sub-complexes: BBsome, IFT-A, and IFT-B. The IFT protein 54 (IFT54) is an important component of the IFT-B sub-complex. In anterograde movement, IFT54 binds to active kinesin-II, walking along the cilia microtubule axoneme and carrying the dynein-2 complex in an inactive state, which works for retrograde movement. Several mutations in IFT54 are known to cause Senior-Loken syndrome, a ciliopathy. IFT54 possesses a divergent Calponin Homology (CH) domain termed as NN-CH domain at its N-terminus. However, several aspects of the function of the NN-CH domain of IFT54 are still obscure. Here, we report the 1H, 15N, and 13C resonance assignments of the NN-CH domain of human IFT54 and its solution structure. The NN-CH domain of human IFT54 adopts essentially the α1–α2–α3–α4–α5 topology as that of mouse IFT54, whose structure was determined by X-ray crystallographic study. The structural information and assignments obtained in this study shed light on the molecular function of the NN-CH domain in IFT54.
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Data availability
The chemical shift assignments for the NN-CH domain of human IFT54 were deposited in the BMRB database under accession number 36589. The atomic coordinates for the ensemble of 20 NMR structures calculated by CYANA 2.1 were deposited in the Protein Data Bank (PDB) under the accession code 2EQO and those with Amber12 refinement under accession code 8KCQ.
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Acknowledgements
We thank Dr. Sumio Sugano of the Department of Medical Genome Sciences, Graduate School of Frontier Sciences, University of Tokyo, for providing the cDNA clone of the human IFT54 NN-CH domain.
Funding
This work was supported by the RIKEN Structural Genomics/Proteomics Initiative (RSGI) and the National Project on Protein Structural and Functional Analyses of the Ministry of Education, Culture, Sports, Science and Technology of Japan. This work was also supported by Gakuin Tokubetsu Kenkyuhi grants from Musashino University to YM.
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A. T., T. K., and M. S. prepared protein samples. K. K., W. D., M. T., K. T., F. H., T. N., N. K., and P. G. contributed to NMR experiments and structure determination. K. K. and Y. M. wrote the manuscript. Y. M. and S. Y. supervised the study.
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Kuwasako, K., Dang, W., He, F. et al. 1H, 13C, and 15N resonance assignments and solution structure of the N-terminal divergent calponin homology (NN-CH) domain of human intraflagellar transport protein 54. Biomol NMR Assign (2024). https://doi.org/10.1007/s12104-024-10170-w
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DOI: https://doi.org/10.1007/s12104-024-10170-w