Regulation of Polycystin-1 Function by Calmodulin Binding

Doerr, Nicholas; Wang, Yidi; Kipp, Kevin R.; Benza, Jesse J.; Плетнев, В. З.; Pavlov, Tengis S.; Staruschenko, Alexander; Mohieldin, Ashraf M.; Takahashi, Maki; Nauli, Surya M.; Weimbs, Thomas

doi:10.1371/journal.pone.0161525

Cited by 18 publications

(15 citation statements)

References 65 publications

(113 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To test whether PP1 and calmodulin function together to regulate PC1 trafficking, we generated 2 more mouse full-length PC1 constructs, YFPPC1-S4134D-HA and YFPPC1-S4134D/K4142A/V4143A-HA, to disrupt the calmodulin-PC1 binding or calmodulin-PC1 and PP1-PC1 binding simultaneously. Consistent with a previous study (38), the S4134D mutation did not impair PC1 ciliary localization. Triple point mutations in PC1 significantly reduced its ciliary localization, whereas the efficiency is similar to that of YFPPC1-K4142A/V4143A-HA ( Fig.…”

Section: Mutations In the Newly Identified Cts Impair Ciliary Trafficsupporting

confidence: 92%

“…Several studies have revealed that PP1 can form a complex with calmodulin, and this complex is essential for intracellular trafficking (20,37). Interestingly, the PC1 CTT also contains a calmodulin-binding site overlapping with the G-protein activation domain, which is close to the PP1 docking motif, and point mutation S4144D (mouse equivalent, S4134D) impairs PC1-calmodulin interaction (38). To test whether PP1 and calmodulin function together to regulate PC1 trafficking, we generated 2 more mouse full-length PC1 constructs, YFPPC1-S4134D-HA and YFPPC1-S4134D/K4142A/V4143A-HA, to disrupt the calmodulin-PC1 binding or calmodulin-PC1 and PP1-PC1 binding simultaneously.…”

Section: Mutations In the Newly Identified Cts Impair Ciliary Trafficmentioning

confidence: 99%

See 1 more Smart Citation

Protein phosphatase 1α interacts with a novel ciliary targeting sequence of polycystin‐1 and regulates polycystin‐1 trafficking

Luo

et al. 2019

FASEB j.

View full text Add to dashboard Cite

Autosomal dominant polycystic kidney disease (ADPKD) is the most common genetic disorder causing renal failure. Mutations of polycystic kidney disease 1 (PKD1) account for most ADPKD cases. Defective ciliary localization of polycystin‐1 (PC1), a large integral membrane protein encoded by PKD1, underlies the pathogenesis of a subgroup of patients with ADPKD. However, the mechanisms by which PC1 and other ciliary proteins traffic to the primary cilium remain poorly understood. A ciliary targeting sequence (CTS) that resides in ciliary receptors is considered to function in the process. It has been reported that the VxP motif in the intracellular C‐terminal tail of PC1 functions as a CTS in an ADP ribosylation factor 4 (Arf4)/ArfGAP with SH3 domain, ankyrin repeat and PH domain 1 (ASAP1)–dependent manner. However, other recent studies have revealed that this motif is dispensable for PC1 trafficking to cilia. In this study, we identified a novel CTS consisting of 8 residues (RHKVRFEG) in the PC1 C tail. We found that this motif is sufficient to bind protein phosphatase 1 (PP1)α, a ubiquitously expressed phosphatase in the phosphoprotein phosphatase (PPP) family. Mutations in this CTS motif disrupt binding with PP1α and impair ciliary localization of PC1. Additionally, short hairpin RNA–mediated knockdown of PP1α results in reduced ciliary localization of PC1 and elongated cilia, suggesting a role for PP1α in the regulation of ciliary structure and function.—Luo, C., Wu, M., Su, X., Yu, F., Brautigan, D. L., Chen, J., Zhou, J. Protein phosphatase 1α interacts with a novel ciliary targeting sequence of polycystin‐1 and regulates polycystin‐1 trafficking. FASEB J. 33, 9945–9958 (2019). http://www.fasebj.org

show abstract

Section: Mutations In the Newly Identified Cts Impair Ciliary Trafficsupporting

confidence: 92%

Section: Mutations In the Newly Identified Cts Impair Ciliary Trafficmentioning

confidence: 99%

Protein phosphatase 1α interacts with a novel ciliary targeting sequence of polycystin‐1 and regulates polycystin‐1 trafficking

Luo

et al. 2019

FASEB j.

View full text Add to dashboard Cite

show abstract

“…A pcDNA5-backbone plasmid for stable expression in Flp-In cells was modified from 61 and expresses full-length human PC1 with N-terminal eGFP and C-terminal HA tags. Constructs expressing PC1-CTT with either eGFP, eGFP-3HA or 3HA C-terminal tags were cloned into pcDNA3 and include the following PC1 sequences: PC1 insert amplified from pcDNA4/TO CD16.7-h-PKD1 FLM (4078-4303) (Addgene plasmid #83465 63 ); and the following mouse PC1-CTT inserts with the addition of 5′ Kozak sequences: PC1-CTT (mPC-4119): ACCatggtggagcttttcctgcgtaggcttcggctctggatgggcttcagcaaggtcaaggagttccgccacaaagtccgctttgaaggaatggatccactgccttcccgctcatccaggggctccaagtcatccccagttgtgctcccacctagctcaggctcagaagcttcacacccatccacctcgtccagccaaccagacgggccgagcgccagcttaagccgctcgacgctgaagctggaaccagagccctctcgcctccatgctgtgtttgaaagtctgcttgtccagtttgaccgactcaaccaggccacagaggacgtctaccagctggagcaacaactccagagccttcaaggccatgggcacaatggacctccttcctctccctcccctggctgcttcccaggctctcagccagctttgcccagccgcctttctcgggccagtcaggggctggatcagactgtaggccccaacagggtgtccctgtggcctaataacaaggtccaccccagcagcact; and a truncated mPC1-4148 lacking the putative mitochondrial targeting sequence (MVELFLRRLRLWMGLSKVKEFRHKVRFEG): ACGATGatggatccactgccttcccgctcatccaggggctccaagtcatccccagttgtgctcccacctagctcaggctcagaagcttcacacccatccacctcgtccagccaaccagacgggccgagcgccagcttaagccgctcgacgctgaagctggaaccagagccctctcgcctccatgctgtgtttgaaagtctgcttgtccagtttgaccgactcaaccaggccacagaggacgtctaccagctggagcaacaactccagagccttcaaggccatgggcacaatggacctccttcctctccctcccctggctgcttcccaggctctcagccagctttgcccagccgcctttctcgggccagtcaggggctggatcagactgtaggccccaacagggtgtccctgtggcctaataacaaggtccaccccagcagcact. MTS-eGFP constructs were generated using gBlocks (Integrated DNA Technologies) and included the MTS sequence (or variants) fused to eGFP.…”

Section: Methodsmentioning

confidence: 99%

A cleavage product of Polycystin-1 is a mitochondrial matrix protein that affects mitochondria morphology and function when heterologously expressed

Lin

Kurashige

Liu

et al. 2018

Sci Rep

View full text Add to dashboard Cite

Recent studies have reported intrinsic metabolic reprogramming in Pkd1 knock-out cells, implicating dysregulated cellular metabolism in the pathogenesis of polycystic kidney disease. However, the exact nature of the metabolic changes and their underlying cause remains controversial. We show herein that Pkd1ko/ko renal epithelial cells have impaired fatty acid utilization, abnormal mitochondrial morphology and function, and that mitochondria in kidneys of ADPKD patients have morphological alterations. We further show that a C-terminal cleavage product of polycystin-1 (CTT) translocates to the mitochondria matrix and that expression of CTT in Pkd1ko/ko cells rescues some of the mitochondrial phenotypes. Using Drosophila to model in vivo effects, we find that transgenic expression of mouse CTT results in decreased viability and exercise endurance but increased CO2 production, consistent with altered mitochondrial function. Our results suggest that PC1 may play a direct role in regulating mitochondrial function and cellular metabolism and provide a framework to understand how impaired mitochondrial function could be linked to the regulation of tubular diameter in both physiological and pathological conditions.

show abstract

“…The first is through the activation of PC-2, enabling an extracellular Ca 2+ influx and leading to the activation of Ca 2+ /CaM-dependent pathways. Secondly, the intracellular tail of PC-1 gets cleaved and translocated to the nucleus, where it regulates DNA transcription via genes related to proliferation [ 65 , 66 , 82 , 83 ].…”

Section: Cilia-mediated and Calcium-dependent Biological Processesmentioning

confidence: 99%

Primary Cilia and Calcium Signaling Interactions

Saternos

Ley

AbouAlaiwi

2020

IJMS

View full text Add to dashboard Cite

The calcium ion (Ca2+) is a diverse secondary messenger with a near-ubiquitous role in a vast array of cellular processes. Cilia are present on nearly every cell type in either a motile or non-motile form; motile cilia generate fluid flow needed for a variety of biological processes, such as left–right body patterning during development, while non-motile cilia serve as the signaling powerhouses of the cell, with vital singling receptors localized to their ciliary membranes. Much of the research currently available on Ca2+-dependent cellular actions and primary cilia are tissue-specific processes. However, basic stimuli-sensing pathways, such as mechanosensation, chemosensation, and electrical sensation (electrosensation), are complex processes entangled in many intersecting pathways; an overview of proposed functions involving cilia and Ca2+ interplay will be briefly summarized here. Next, we will focus on summarizing the evidence for their interactions in basic cellular activities, including the cell cycle, cell polarity and migration, neuronal pattering, glucose-mediated insulin secretion, biliary regulation, and bone formation. Literature investigating the role of cilia and Ca2+-dependent processes at a single-cellular level appears to be scarce, though overlapping signaling pathways imply that cilia and Ca2+ interact with each other on this level in widespread and varied ways on a perpetual basis. Vastly different cellular functions across many different cell types depend on context-specific Ca2+ and cilia interactions to trigger the correct physiological responses, and abnormalities in these interactions, whether at the tissue or the single-cell level, can result in diseases known as ciliopathies; due to their clinical relevance, pathological alterations of cilia function and Ca2+ signaling will also be briefly touched upon throughout this review.

show abstract

Regulation of Polycystin-1 Function by Calmodulin Binding

Cited by 18 publications

References 65 publications

Protein phosphatase 1α interacts with a novel ciliary targeting sequence of polycystin‐1 and regulates polycystin‐1 trafficking

Protein phosphatase 1α interacts with a novel ciliary targeting sequence of polycystin‐1 and regulates polycystin‐1 trafficking

A cleavage product of Polycystin-1 is a mitochondrial matrix protein that affects mitochondria morphology and function when heterologously expressed

Primary Cilia and Calcium Signaling Interactions

Contact Info

Product

Resources

About