Format

Send to

Choose Destination
J Cell Physiol. 2018 Oct;233(10):6877-6895. doi: 10.1002/jcp.26578. Epub 2018 Apr 25.

Tctex-1, a novel interaction partner of Kidney Injury Molecule-1, is required for efferocytosis.

Author information

1
Matthew Mailing Center for Translational Transplant Studies, London Health Sciences Center, Lawson Health Research Institute, London, Ontario, Canada.
2
Renal Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.
3
Burnett School of Biomedical Sciences, University of Central Florida College of Medicine, Orlando, Florida.
4
Division of Nephrology, Department of Medicine, Schulich School of Medicine and Dentistry, London, Western University, Ontario, Canada.

Abstract

Kidney injury molecule-1 (KIM-1) is a phosphatidylserine receptor that is specifically upregulated on proximal tubular epithelial cells (PTECs) during acute kidney injury and mitigates tissue damage by mediating efferocytosis (the phagocytic clearance of apoptotic cells). The signaling molecules that regulate efferocytosis in TECs are not well understood. Using a yeast two-hybrid screen, we identified the dynein light chain protein, Tctex-1, as a novel KIM-1-interacting protein. Immunoprecipitation and confocal imaging studies suggested that Tctex-1 associates with KIM-1 in cells at baseline, but, dissociates from KIM-1 within 90 min of initiation of efferocytosis. Interfering with actin or microtubule polymerization interestingly prevented the dissociation of KIM-1 from Tctex-1. Moreover, the subcellular localization of Tctex-1 changed from being microtubule-associated to mainly cytosolic upon expression of KIM-1. Short hairpin RNA-mediated silencing of endogenous Tctex-1 in cells significantly inhibited efferocytosis to levels comparable to that of knock down of KIM-1 in the same cells. Importantly, Tctex-1 was not involved in the delivery of KIM-1 to the cell-surface. On the other hand, KIM-1 expression significantly inhibited the phosphorylation of Tctex-1 at threonine 94 (T94), a post-translational modification which is known to disrupt the binding of Tctex-1 to dynein on microtubules. In keeping with this, we found that KIM-1 bound less efficiently to the phosphomimic (T94E) mutant of Tctex-1 compared to wild type Tctex-1. Surprisingly, expression of Tctex-1 T94E did not influence KIM-1-mediated efferocytosis. Our studies uncover a previously unknown role for Tctex-1 in KIM-1-dependent efferocytosis in epithelial cells.

KEYWORDS:

cytoskeleton; efferocytosis; kidney injury molecule-1; phagocytosis; tctex-1

PMID:
29693725
PMCID:
PMC6066178
[Available on 2019-10-01]
DOI:
10.1002/jcp.26578

Supplemental Content

Full text links

Icon for Wiley
Loading ...
Support Center