Document Detail


Muscle-specific f-box only proteins facilitate bk channel β(1) subunit downregulation in vascular smooth muscle cells of diabetes mellitus.
MedLine Citation:
PMID:  20966391     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
RATIONALE: activity of the large conductance Ca(2+)-activated K(+) (BK) channels is profoundly modulated by its β(1) subunit (BK-β(1)). However, BK-β(1) expression is downregulated in diabetic vessels. The ubiquitin-proteasome system (UPS) is a major mechanism of intracellular protein degradation. Whether UPS participates in BK-β(1) downregulation in diabetic vessels is unknown.
OBJECTIVE: we hypothesize that UPS facilitates vascular BK-β(1) degradation in diabetes.
METHODS AND RESULTS: using patch clamp and molecular biological approaches, we found that BK-β(1)-mediated channel activation and BK-β(1) protein expression were reduced in aortas of streptozotocin-induced diabetic rats and in human coronary arterial smooth muscle cells (CASMCs) cultured in high glucose. This was accompanied by upregulation of F-box only protein (FBXO)-9 and FBXO-32 (atrogin-1), the key components of the Skp1-Cullin-F-box (SCF) type ubiquitin ligase complex. BK-β(1) expression was suppressed by the FBXO activator doxorubicin but enhanced by FBXO-9 small interfering RNA or by the proteasome inhibitor MG-132. Cotransfection of atrogin-1 in HEK293 cells significantly reduced Flag-hSlo-β(1) expression by 2.16-fold, compared with expression of Flag-hSlo-β(1)V146A (a mutant without the PDZ-binding motif). After cotransfection with atrogin-1, the ubiquitination of Flag-hSlo-β(1) was increased by 1.91-fold, compared with that of hSlo-β(1)V146A, whereas cotransfection with atrogin-1ΔF (a nonfunctional mutant without the F-box motif) had no effect. Moreover, inhibition of Akt signaling attenuated the phosphorylation of forkhead box O transcription factor (FOXO)-3a and enhanced atrogin-1 expression, which in turn suppressed BK-β(1) protein levels in human CASMCs.
CONCLUSIONS: downregulation of vascular BK-β(1) expression in diabetes and in high-glucose culture conditions was associated with FOXO-3a/FBXO-dependent increase in BK-β(1) degradation.
Authors:
Dai-Min Zhang; Tongrong He; Zvonimir S Katusic; Hon-Chi Lee; Tong Lu
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-10-21
Journal Detail:
Title:  Circulation research     Volume:  107     ISSN:  1524-4571     ISO Abbreviation:  Circ. Res.     Publication Date:  2010 Dec 
Date Detail:
Created Date:  2010-12-14     Completed Date:  2011-01-24     Revised Date:  2012-04-11    
Medline Journal Info:
Nlm Unique ID:  0047103     Medline TA:  Circ Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1454-9     Citation Subset:  IM    
Affiliation:
Division of Cardiovascular Diseases, Department of Internal Medicine, Mayo Clinic, 200 First St SW, Rochester, MN 55905, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Cells, Cultured
Diabetes Mellitus / metabolism*
Down-Regulation*
F-Box Proteins / physiology*
Forkhead Transcription Factors / physiology
Glucose
Humans
Large-Conductance Calcium-Activated Potassium Channels / metabolism*
Muscle, Smooth, Vascular / pathology*
Myocytes, Smooth Muscle / metabolism*
Proteasome Endopeptidase Complex / metabolism
Protein Stability
Protein Subunits / metabolism
Ubiquitin / metabolism
Grant Support
ID/Acronym/Agency:
HL080118/HL/NHLBI NIH HHS; HL74180/HL/NHLBI NIH HHS; HL91867/HL/NHLBI NIH HHS; R01 HL074180-04/HL/NHLBI NIH HHS; R01 HL074180-08/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/F-Box Proteins; 0/FOXO3 protein, human; 0/Forkhead Transcription Factors; 0/Large-Conductance Calcium-Activated Potassium Channels; 0/Protein Subunits; 0/Ubiquitin; 50-99-7/Glucose; EC 3.4.25.1/Proteasome Endopeptidase Complex

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Passive participle marking by african american english-speaking children reared in poverty.
Next Document:  In the RyR2(R4496C) mouse model of CPVT, ?-adrenergic stimulation induces Ca waves by increasing SR ...