Enhanced ubiquitination of cytoskeletal proteins in pressure overloaded myocardium is accompanied by changes in specific E3 ligases

Sundaravadivel Balasubramanian, Santhoshkumar Mani, Hirokazu Shiraishi, Rebecca K. Johnston, Kentaro Yamane, Christopher D. Willey, George Cooper IV, William J. Tuxworth, Dhandapani Kuppuswamy

Research output: Contribution to journalArticle

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Abstract

Ubiquitin conjugation of proteins is critical for cell homeostasis and contributes to both cell survival and death. Here we studied ubiquitination of proteins in pressure overloaded (PO) myocardium in the context of cardiomyocyte survival. Analysis using a feline right ventricular pressure overload (RVPO) model revealed a robust and transient increase in ubiquitination of proteins present in the Triton X-100-insoluble fraction in 24 to 48 h PO myocardium, and confocal micrographs indicate this increase in ubiquitination occurs subsarcolemmaly near the intercalated disc area of cardiomyocytes. The ubiquitination was accompanied by changes in E3 ligases including Cbl, E6AP, Mdm2 and cIAP in the same period of PO, although atrophy-related E3 ligases, MuRF1 and MuRF3 were unaltered. Furthermore, Cbl displayed a substantial increase in both levels of expression and tyrosine phosphorylation in 48 h PO myocardium. Confocal studies revealed enrichment of Cbl at the intercalated discs of 48 h PO cardiomyocytes, as evidenced by its colocalization with N-cadherin. Although apoptosis was observed in 48 h PO myocardium by TUNEL staining, cardiomyocytes showing ubiquitin staining were not positive for TUNEL staining. Furthermore, 48 h PO resulted in the phosphorylation of inhibitor of nuclear factor kappa B (IκB), suggesting its ubiquitin-mediated degradation and the nuclear localization of NFκB for the expression of specific cell survival factors such as cIAPs. Together these data indicate that increased levels of E3 ligases that regulate cell homeostasis and promote cell survival could ubiquitinate multiple cytoskeletal protein targets and that these events that occur during the early phase of PO may contribute to both cardiomyocyte survival and hypertrophy.

Original languageEnglish (US)
Pages (from-to)669-679
Number of pages11
JournalJournal of Molecular and Cellular Cardiology
Volume41
Issue number4
DOIs
StatePublished - Oct 1 2006

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Ubiquitin-Protein Ligases
Cytoskeletal Proteins
Ubiquitination
Myocardium
Pressure
Cardiac Myocytes
Ubiquitin
Cell Survival
In Situ Nick-End Labeling
Staining and Labeling
Homeostasis
Phosphorylation
Proteins
NF-kappa B
Felidae
Octoxynol
Ventricular Pressure
Cadherins
Hypertrophy
Atrophy

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

Balasubramanian, Sundaravadivel ; Mani, Santhoshkumar ; Shiraishi, Hirokazu ; Johnston, Rebecca K. ; Yamane, Kentaro ; Willey, Christopher D. ; Cooper IV, George ; Tuxworth, William J. ; Kuppuswamy, Dhandapani. / Enhanced ubiquitination of cytoskeletal proteins in pressure overloaded myocardium is accompanied by changes in specific E3 ligases. In: Journal of Molecular and Cellular Cardiology. 2006 ; Vol. 41, No. 4. pp. 669-679.
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abstract = "Ubiquitin conjugation of proteins is critical for cell homeostasis and contributes to both cell survival and death. Here we studied ubiquitination of proteins in pressure overloaded (PO) myocardium in the context of cardiomyocyte survival. Analysis using a feline right ventricular pressure overload (RVPO) model revealed a robust and transient increase in ubiquitination of proteins present in the Triton X-100-insoluble fraction in 24 to 48 h PO myocardium, and confocal micrographs indicate this increase in ubiquitination occurs subsarcolemmaly near the intercalated disc area of cardiomyocytes. The ubiquitination was accompanied by changes in E3 ligases including Cbl, E6AP, Mdm2 and cIAP in the same period of PO, although atrophy-related E3 ligases, MuRF1 and MuRF3 were unaltered. Furthermore, Cbl displayed a substantial increase in both levels of expression and tyrosine phosphorylation in 48 h PO myocardium. Confocal studies revealed enrichment of Cbl at the intercalated discs of 48 h PO cardiomyocytes, as evidenced by its colocalization with N-cadherin. Although apoptosis was observed in 48 h PO myocardium by TUNEL staining, cardiomyocytes showing ubiquitin staining were not positive for TUNEL staining. Furthermore, 48 h PO resulted in the phosphorylation of inhibitor of nuclear factor kappa B (IκB), suggesting its ubiquitin-mediated degradation and the nuclear localization of NFκB for the expression of specific cell survival factors such as cIAPs. Together these data indicate that increased levels of E3 ligases that regulate cell homeostasis and promote cell survival could ubiquitinate multiple cytoskeletal protein targets and that these events that occur during the early phase of PO may contribute to both cardiomyocyte survival and hypertrophy.",
author = "Sundaravadivel Balasubramanian and Santhoshkumar Mani and Hirokazu Shiraishi and Johnston, {Rebecca K.} and Kentaro Yamane and Willey, {Christopher D.} and {Cooper IV}, George and Tuxworth, {William J.} and Dhandapani Kuppuswamy",
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Balasubramanian, S, Mani, S, Shiraishi, H, Johnston, RK, Yamane, K, Willey, CD, Cooper IV, G, Tuxworth, WJ & Kuppuswamy, D 2006, 'Enhanced ubiquitination of cytoskeletal proteins in pressure overloaded myocardium is accompanied by changes in specific E3 ligases', Journal of Molecular and Cellular Cardiology, vol. 41, no. 4, pp. 669-679. https://doi.org/10.1016/j.yjmcc.2006.04.022

Enhanced ubiquitination of cytoskeletal proteins in pressure overloaded myocardium is accompanied by changes in specific E3 ligases. / Balasubramanian, Sundaravadivel; Mani, Santhoshkumar; Shiraishi, Hirokazu; Johnston, Rebecca K.; Yamane, Kentaro; Willey, Christopher D.; Cooper IV, George; Tuxworth, William J.; Kuppuswamy, Dhandapani.

In: Journal of Molecular and Cellular Cardiology, Vol. 41, No. 4, 01.10.2006, p. 669-679.

Research output: Contribution to journalArticle

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T1 - Enhanced ubiquitination of cytoskeletal proteins in pressure overloaded myocardium is accompanied by changes in specific E3 ligases

AU - Balasubramanian, Sundaravadivel

AU - Mani, Santhoshkumar

AU - Shiraishi, Hirokazu

AU - Johnston, Rebecca K.

AU - Yamane, Kentaro

AU - Willey, Christopher D.

AU - Cooper IV, George

AU - Tuxworth, William J.

AU - Kuppuswamy, Dhandapani

PY - 2006/10/1

Y1 - 2006/10/1

N2 - Ubiquitin conjugation of proteins is critical for cell homeostasis and contributes to both cell survival and death. Here we studied ubiquitination of proteins in pressure overloaded (PO) myocardium in the context of cardiomyocyte survival. Analysis using a feline right ventricular pressure overload (RVPO) model revealed a robust and transient increase in ubiquitination of proteins present in the Triton X-100-insoluble fraction in 24 to 48 h PO myocardium, and confocal micrographs indicate this increase in ubiquitination occurs subsarcolemmaly near the intercalated disc area of cardiomyocytes. The ubiquitination was accompanied by changes in E3 ligases including Cbl, E6AP, Mdm2 and cIAP in the same period of PO, although atrophy-related E3 ligases, MuRF1 and MuRF3 were unaltered. Furthermore, Cbl displayed a substantial increase in both levels of expression and tyrosine phosphorylation in 48 h PO myocardium. Confocal studies revealed enrichment of Cbl at the intercalated discs of 48 h PO cardiomyocytes, as evidenced by its colocalization with N-cadherin. Although apoptosis was observed in 48 h PO myocardium by TUNEL staining, cardiomyocytes showing ubiquitin staining were not positive for TUNEL staining. Furthermore, 48 h PO resulted in the phosphorylation of inhibitor of nuclear factor kappa B (IκB), suggesting its ubiquitin-mediated degradation and the nuclear localization of NFκB for the expression of specific cell survival factors such as cIAPs. Together these data indicate that increased levels of E3 ligases that regulate cell homeostasis and promote cell survival could ubiquitinate multiple cytoskeletal protein targets and that these events that occur during the early phase of PO may contribute to both cardiomyocyte survival and hypertrophy.

AB - Ubiquitin conjugation of proteins is critical for cell homeostasis and contributes to both cell survival and death. Here we studied ubiquitination of proteins in pressure overloaded (PO) myocardium in the context of cardiomyocyte survival. Analysis using a feline right ventricular pressure overload (RVPO) model revealed a robust and transient increase in ubiquitination of proteins present in the Triton X-100-insoluble fraction in 24 to 48 h PO myocardium, and confocal micrographs indicate this increase in ubiquitination occurs subsarcolemmaly near the intercalated disc area of cardiomyocytes. The ubiquitination was accompanied by changes in E3 ligases including Cbl, E6AP, Mdm2 and cIAP in the same period of PO, although atrophy-related E3 ligases, MuRF1 and MuRF3 were unaltered. Furthermore, Cbl displayed a substantial increase in both levels of expression and tyrosine phosphorylation in 48 h PO myocardium. Confocal studies revealed enrichment of Cbl at the intercalated discs of 48 h PO cardiomyocytes, as evidenced by its colocalization with N-cadherin. Although apoptosis was observed in 48 h PO myocardium by TUNEL staining, cardiomyocytes showing ubiquitin staining were not positive for TUNEL staining. Furthermore, 48 h PO resulted in the phosphorylation of inhibitor of nuclear factor kappa B (IκB), suggesting its ubiquitin-mediated degradation and the nuclear localization of NFκB for the expression of specific cell survival factors such as cIAPs. Together these data indicate that increased levels of E3 ligases that regulate cell homeostasis and promote cell survival could ubiquitinate multiple cytoskeletal protein targets and that these events that occur during the early phase of PO may contribute to both cardiomyocyte survival and hypertrophy.

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