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HSC70 is a chaperone for wild-type and mutant cardiac myosin binding protein C

Cardiac myosin binding protein C (MYBPC3) is the most commonly mutated gene associated with hypertrophic cardiomyopathy (HCM). Haploinsufficiency of full-length MYBPC3 and disruption of proteostasis have both been proposed as central to HCM disease pathogenesis. Discriminating the relative contribut...

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Published in:JCI insight 2018-06, Vol.3 (11)
Main Authors: Glazier, Amelia A, Hafeez, Neha, Mellacheruvu, Dattatreya, Basrur, Venkatesha, Nesvizhskii, Alexey I, Lee, Lap Man, Shao, Hao, Tang, Vi, Yob, Jaime M, Gestwicki, Jason E, Helms, Adam S, Day, Sharlene M
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cited_by cdi_FETCH-LOGICAL-c465t-f828a498bd7b656412eee8cc74ae0e30735fddbd90ef902b603c0c580f8a48e93
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container_issue 11
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container_title JCI insight
container_volume 3
creator Glazier, Amelia A
Hafeez, Neha
Mellacheruvu, Dattatreya
Basrur, Venkatesha
Nesvizhskii, Alexey I
Lee, Lap Man
Shao, Hao
Tang, Vi
Yob, Jaime M
Gestwicki, Jason E
Helms, Adam S
Day, Sharlene M
description Cardiac myosin binding protein C (MYBPC3) is the most commonly mutated gene associated with hypertrophic cardiomyopathy (HCM). Haploinsufficiency of full-length MYBPC3 and disruption of proteostasis have both been proposed as central to HCM disease pathogenesis. Discriminating the relative contributions of these 2 mechanisms requires fundamental knowledge of how turnover of WT and mutant MYBPC3 proteins is regulated. We expressed several disease-causing mutations in MYBPC3 in primary neonatal rat ventricular cardiomyocytes. In contrast to WT MYBPC3, mutant proteins showed reduced expression and failed to localize to the sarcomere. In an unbiased coimmunoprecipitation/mass spectrometry screen, we identified HSP70-family chaperones as interactors of both WT and mutant MYBPC3. Heat shock cognate 70 kDa (HSC70) was the most abundant chaperone interactor. Knockdown of HSC70 significantly slowed degradation of both WT and mutant MYBPC3, while pharmacologic activation of HSC70 and HSP70 accelerated degradation. HSC70 was expressed in discrete striations in the sarcomere. Expression of mutant MYBPC3 did not affect HSC70 localization, nor did it induce a protein folding stress response or ubiquitin proteasome dysfunction. Together these data suggest that WT and mutant MYBPC3 proteins are clients for HSC70, and that the HSC70 chaperone system plays a major role in regulating MYBPC3 protein turnover.
doi_str_mv 10.1172/jci.insight.99319
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Haploinsufficiency of full-length MYBPC3 and disruption of proteostasis have both been proposed as central to HCM disease pathogenesis. Discriminating the relative contributions of these 2 mechanisms requires fundamental knowledge of how turnover of WT and mutant MYBPC3 proteins is regulated. We expressed several disease-causing mutations in MYBPC3 in primary neonatal rat ventricular cardiomyocytes. In contrast to WT MYBPC3, mutant proteins showed reduced expression and failed to localize to the sarcomere. In an unbiased coimmunoprecipitation/mass spectrometry screen, we identified HSP70-family chaperones as interactors of both WT and mutant MYBPC3. Heat shock cognate 70 kDa (HSC70) was the most abundant chaperone interactor. Knockdown of HSC70 significantly slowed degradation of both WT and mutant MYBPC3, while pharmacologic activation of HSC70 and HSP70 accelerated degradation. HSC70 was expressed in discrete striations in the sarcomere. Expression of mutant MYBPC3 did not affect HSC70 localization, nor did it induce a protein folding stress response or ubiquitin proteasome dysfunction. Together these data suggest that WT and mutant MYBPC3 proteins are clients for HSC70, and that the HSC70 chaperone system plays a major role in regulating MYBPC3 protein turnover.</abstract><cop>United States</cop><pub>American Society for Clinical Investigation</pub><pmid>29875314</pmid><doi>10.1172/jci.insight.99319</doi><orcidid>https://orcid.org/0000-0003-2506-4329</orcidid><orcidid>https://orcid.org/0000-0001-6079-9756</orcidid><orcidid>https://orcid.org/0000-0001-8221-7673</orcidid><oa>free_for_read</oa></addata></record>
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subjects Acetylcysteine - analogs & derivatives
Acetylcysteine - pharmacology
Animals
Animals, Newborn
Cardiomyopathy, Hypertrophic - genetics
Cardiomyopathy, Hypertrophic - pathology
Carrier Proteins - genetics
Carrier Proteins - metabolism
Cell Nucleus - metabolism
Gene Knockdown Techniques
Haploinsufficiency
HEK293 Cells
HSC70 Heat-Shock Proteins - genetics
HSC70 Heat-Shock Proteins - metabolism
Humans
Myocardium - pathology
Proteasome Endopeptidase Complex - drug effects
Proteasome Inhibitors - pharmacology
Proteolysis - drug effects
Proteostasis - genetics
Rats
Sarcomeres - pathology
Ventricular Septum - pathology
title HSC70 is a chaperone for wild-type and mutant cardiac myosin binding protein C
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