Supervillin couples myosin-dependent contractility to podosomes and enables their turnover.
Standard
Supervillin couples myosin-dependent contractility to podosomes and enables their turnover. / Bhuwania, Rahidmina; Cornfine, Susanne; Fang, Zhiyou; Krüger, Marcus; Luna, Elizabeth J; Linder, Stefan.
in: J CELL SCI, Jahrgang 125, Nr. Pt 9, Pt 9, 2012, S. 2300-2314.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Supervillin couples myosin-dependent contractility to podosomes and enables their turnover.
AU - Bhuwania, Rahidmina
AU - Cornfine, Susanne
AU - Fang, Zhiyou
AU - Krüger, Marcus
AU - Luna, Elizabeth J
AU - Linder, Stefan
PY - 2012
Y1 - 2012
N2 - Podosomes are actin-rich adhesion and invasion structures. Especially in macrophages, podosomes exist in two subpopulations, large precursors at the cell periphery and smaller podosomes (successors) in the cell interior. To date, the mechanisms that differentially regulate these subpopulations are largely unknown. Here, we show that the membrane-associated protein supervillin localizes preferentially to successor podosomes and becomes enriched at precursors immediately before their dissolution. Consistently, podosome numbers are inversely correlated with supervillin protein levels. Using deletion constructs, we find that the myosin II regulatory N-terminus of supervillin [SV(1-174)] is crucial for these effects. Phosphorylated myosin light chain (pMLC) localizes at supervillin-positive podosomes, and time-lapse analyses show that enrichment of GFP-supervillin at podosomes coincides with their coupling to contractile myosin-IIA-positive cables. We also show that supervillin binds only to activated myosin IIA, and a dysregulated N-terminal construct [SV(1-830)] enhances pMLC levels at podosomes. Thus, preferential recruitment of supervillin to podosome subpopulations might both require and induce actomyosin contractility. Using siRNA and pharmacological inhibition, we demonstrate that supervillin and myosin IIA cooperate to regulate podosome lifetime, podosomal matrix degradation and cell polarization. In sum, we show here that podosome subpopulations differ in their molecular composition and identify supervillin, in cooperation with myosin IIA, as a crucial factor in the regulation of podosome turnover and function.
AB - Podosomes are actin-rich adhesion and invasion structures. Especially in macrophages, podosomes exist in two subpopulations, large precursors at the cell periphery and smaller podosomes (successors) in the cell interior. To date, the mechanisms that differentially regulate these subpopulations are largely unknown. Here, we show that the membrane-associated protein supervillin localizes preferentially to successor podosomes and becomes enriched at precursors immediately before their dissolution. Consistently, podosome numbers are inversely correlated with supervillin protein levels. Using deletion constructs, we find that the myosin II regulatory N-terminus of supervillin [SV(1-174)] is crucial for these effects. Phosphorylated myosin light chain (pMLC) localizes at supervillin-positive podosomes, and time-lapse analyses show that enrichment of GFP-supervillin at podosomes coincides with their coupling to contractile myosin-IIA-positive cables. We also show that supervillin binds only to activated myosin IIA, and a dysregulated N-terminal construct [SV(1-830)] enhances pMLC levels at podosomes. Thus, preferential recruitment of supervillin to podosome subpopulations might both require and induce actomyosin contractility. Using siRNA and pharmacological inhibition, we demonstrate that supervillin and myosin IIA cooperate to regulate podosome lifetime, podosomal matrix degradation and cell polarization. In sum, we show here that podosome subpopulations differ in their molecular composition and identify supervillin, in cooperation with myosin IIA, as a crucial factor in the regulation of podosome turnover and function.
KW - Humans
KW - Protein Structure, Tertiary
KW - Phosphorylation
KW - Transfection
KW - Protein Binding
KW - Plasmids
KW - Actins/metabolism
KW - RNA, Small Interfering
KW - Actomyosin/metabolism
KW - Cell Polarity/physiology
KW - Cell Surface Extensions/physiology/ultrastructure
KW - Membrane Proteins/chemistry/genetics/metabolism
KW - Microfilament Proteins/chemistry/genetics/metabolism
KW - Monocytes/cytology/physiology/ultrastructure
KW - Myosin Light Chains/genetics/metabolism
KW - Nonmuscle Myosin Type IIA/genetics/metabolism
KW - Time-Lapse Imaging
KW - Humans
KW - Protein Structure, Tertiary
KW - Phosphorylation
KW - Transfection
KW - Protein Binding
KW - Plasmids
KW - Actins/metabolism
KW - RNA, Small Interfering
KW - Actomyosin/metabolism
KW - Cell Polarity/physiology
KW - Cell Surface Extensions/physiology/ultrastructure
KW - Membrane Proteins/chemistry/genetics/metabolism
KW - Microfilament Proteins/chemistry/genetics/metabolism
KW - Monocytes/cytology/physiology/ultrastructure
KW - Myosin Light Chains/genetics/metabolism
KW - Nonmuscle Myosin Type IIA/genetics/metabolism
KW - Time-Lapse Imaging
M3 - SCORING: Journal article
VL - 125
SP - 2300
EP - 2314
JO - J CELL SCI
JF - J CELL SCI
SN - 0021-9533
IS - Pt 9
M1 - Pt 9
ER -
