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Structural definition of the F-actin–binding THATCH domain from HIP1R

Nature Structural & Molecular Biology volume 13, pages 121–130 (2006)Cite this article

Abstract

Huntingtin-interacting protein-1 related (HIP1R) has a crucial protein-trafficking role, mediating associations between actin and clathrin-coated structures at the plasma membrane and trans-Golgi network. Here, we characterize the F-actin–binding region of HIP1R, termed the talin-HIP1/R/Sla2p actin-tethering C-terminal homology (THATCH) domain. The 1.9-Å crystal structure of the human HIP1R THATCH core reveals a large sequence-conserved surface patch created primarily by residues from the third and fourth helices of a unique five-helix bundle. Point mutations of seven contiguous patch residues produced significant decreases in F-actin binding. We also show that THATCH domains have a conserved C-terminal latch capable of oligomerizing the core, thereby modulating F-actin engagement. Collectively, these results establish a framework for investigating the links between endocytosis and actin dynamics mediated by THATCH domain–containing proteins.

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Figure 1: Sequence conservation and occurrence of THATCH domains.
Figure 2: Crystal structure of the HIP1R THATCH domain core.
Figure 3: HIP1R THATCH core binds F-actin to a lesser extent than HIP1R THATCH owing to loss of oligomerization through the THATCH domain latch.
Figure 4: Surface analysis of the HIP1R THATCH core reveals a potential F-actin–binding surface.
Figure 5: Mutational analysis of the HIP1R THATCH domain reveals an F-actin–binding surface.

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Acknowledgements

We thank J. Heuser for helpful discussions regarding his deep-etch EM studies of HIP1R, J. Alexander-Brett for assistance with SPR data analysis, L. Traub and C. Nelson for critical comments on the manuscript and Z. Yang and J. Philie for technical assistance. This work was supported by US National Institutes of Health grant GM62414-04 (Midwest Center for Structural Genomics, to D.H.F.; ID code APC35300) and by Canadian Institutes of Health Research grant MOP-15396 (to P.S.M.).

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Authors and Affiliations

  1. Department of Pathology and Immunology, Washington University School of Medicine, 660 S. Euclid Ave, St. Louis, 63110, Missouri, USA

    Tom J Brett & Daved H Fremont

  2. Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 660 S. Euclid Ave, St. Louis, 63110, Missouri, USA

    Daved H Fremont

  3. Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, 3801 University Street, Montreal, H3A 2B4, Quebec, Canada

    Valerie Legendre-Guillemin & Peter S McPherson

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  1. Tom J Brett
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Correspondence to Daved H Fremont.

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Brett, T., Legendre-Guillemin, V., McPherson, P. et al. Structural definition of the F-actin–binding THATCH domain from HIP1R. Nat Struct Mol Biol 13, 121–130 (2006). https://doi.org/10.1038/nsmb1043

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