Live-cell single-molecule imaging reveals clathrin and caveolin-1 dependent docking of SMAD4 at the cell membrane

Yong Yang; Joy Wolfram; Jianliang Shen; Yuliang Zhao; Xiaohong Fang; Haifa Shen; Mauro Ferrari

doi:10.1016/j.febslet.2013.10.041

Live-cell single-molecule imaging reveals clathrin and caveolin-1 dependent docking of SMAD4 at the cell membrane

Yong Yang, Joy Wolfram, Jianliang Shen, Yuliang Zhao, Xiaohong Fang, Haifa Shen, Mauro Ferrari

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

Transforming growth factor β (TGF-β) signaling is important for many biological processes. Although the sequential events of this cascade are known, the dynamics remain speculative. Here, live-cell single-molecule total internal reflection fluorescence microscopy was used to monitor the dynamics of SMAD4, a TGF-β downstream effector, in MDA-MB-231 breast cancer cells. Contrary to previous belief, SMAD4 was detectable at the cytoplasmic membrane, displaying two subpopulations with different membrane docking behaviors. These subpopulations were regulated by clathrin and caveolin-1, and had opposing roles in the nuclear shuttling of SMAD4 and the subsequent transcriptional regulation of genes associated with cell migration. The notion that membrane-docking behaviors of downstream molecules could predict the cellular response to growth factors may revolutionize the way we view cell signaling.

Original language	English (US)
Pages (from-to)	3912-3920
Number of pages	9
Journal	FEBS Letters
Volume	587
Issue number	24
DOIs	https://doi.org/10.1016/j.febslet.2013.10.041
State	Published - Dec 11 2013

Keywords

Caveolin-1
Clathrin
Membrane docking
Single-molecule fluorescence imaging
SMAD4

ASJC Scopus subject areas

Biochemistry
Biophysics
Cell Biology
Genetics
Molecular Biology
Structural Biology

Access to Document

10.1016/j.febslet.2013.10.041

Cite this

@article{afd59e1073d247f18b3f98d75c0e500c,

title = "Live-cell single-molecule imaging reveals clathrin and caveolin-1 dependent docking of SMAD4 at the cell membrane",

abstract = "Transforming growth factor β (TGF-β) signaling is important for many biological processes. Although the sequential events of this cascade are known, the dynamics remain speculative. Here, live-cell single-molecule total internal reflection fluorescence microscopy was used to monitor the dynamics of SMAD4, a TGF-β downstream effector, in MDA-MB-231 breast cancer cells. Contrary to previous belief, SMAD4 was detectable at the cytoplasmic membrane, displaying two subpopulations with different membrane docking behaviors. These subpopulations were regulated by clathrin and caveolin-1, and had opposing roles in the nuclear shuttling of SMAD4 and the subsequent transcriptional regulation of genes associated with cell migration. The notion that membrane-docking behaviors of downstream molecules could predict the cellular response to growth factors may revolutionize the way we view cell signaling.",

keywords = "Caveolin-1, Clathrin, Membrane docking, Single-molecule fluorescence imaging, SMAD4",

author = "Yong Yang and Joy Wolfram and Jianliang Shen and Yuliang Zhao and Xiaohong Fang and Haifa Shen and Mauro Ferrari",

note = "Funding Information: We are very grateful for the guidance of Dr. Joan Massagu{\'e} and Dr. Laurel Raftery during the preparation of this manuscript. This research was supported by funds from the Methodist Hospital Research Institute. Partial funds were from the following sources: the Ernest Cockrell Jr. Distinguished Endowed Chair (M.F.), the US Department of Defense ( W81XWH-09-1-0212 ) (M.F.), the National Institute of Health ( U54CA143837 , U54CA151668 ) (M.F.), Nylands nation Finland (J.W.), Department of Defense grant W81XWH-12-1-0414 (M.F.) and the State of Texas CPRIT grant RP121071 (M.F. and H.S.). ",

year = "2013",

month = dec,

day = "11",

doi = "10.1016/j.febslet.2013.10.041",

language = "English (US)",

volume = "587",

pages = "3912--3920",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "Elsevier",

number = "24",

}

TY - JOUR

T1 - Live-cell single-molecule imaging reveals clathrin and caveolin-1 dependent docking of SMAD4 at the cell membrane

AU - Yang, Yong

AU - Wolfram, Joy

AU - Shen, Jianliang

AU - Zhao, Yuliang

AU - Fang, Xiaohong

AU - Shen, Haifa

AU - Ferrari, Mauro

N1 - Funding Information: We are very grateful for the guidance of Dr. Joan Massagué and Dr. Laurel Raftery during the preparation of this manuscript. This research was supported by funds from the Methodist Hospital Research Institute. Partial funds were from the following sources: the Ernest Cockrell Jr. Distinguished Endowed Chair (M.F.), the US Department of Defense ( W81XWH-09-1-0212 ) (M.F.), the National Institute of Health ( U54CA143837 , U54CA151668 ) (M.F.), Nylands nation Finland (J.W.), Department of Defense grant W81XWH-12-1-0414 (M.F.) and the State of Texas CPRIT grant RP121071 (M.F. and H.S.).

PY - 2013/12/11

Y1 - 2013/12/11

N2 - Transforming growth factor β (TGF-β) signaling is important for many biological processes. Although the sequential events of this cascade are known, the dynamics remain speculative. Here, live-cell single-molecule total internal reflection fluorescence microscopy was used to monitor the dynamics of SMAD4, a TGF-β downstream effector, in MDA-MB-231 breast cancer cells. Contrary to previous belief, SMAD4 was detectable at the cytoplasmic membrane, displaying two subpopulations with different membrane docking behaviors. These subpopulations were regulated by clathrin and caveolin-1, and had opposing roles in the nuclear shuttling of SMAD4 and the subsequent transcriptional regulation of genes associated with cell migration. The notion that membrane-docking behaviors of downstream molecules could predict the cellular response to growth factors may revolutionize the way we view cell signaling.

AB - Transforming growth factor β (TGF-β) signaling is important for many biological processes. Although the sequential events of this cascade are known, the dynamics remain speculative. Here, live-cell single-molecule total internal reflection fluorescence microscopy was used to monitor the dynamics of SMAD4, a TGF-β downstream effector, in MDA-MB-231 breast cancer cells. Contrary to previous belief, SMAD4 was detectable at the cytoplasmic membrane, displaying two subpopulations with different membrane docking behaviors. These subpopulations were regulated by clathrin and caveolin-1, and had opposing roles in the nuclear shuttling of SMAD4 and the subsequent transcriptional regulation of genes associated with cell migration. The notion that membrane-docking behaviors of downstream molecules could predict the cellular response to growth factors may revolutionize the way we view cell signaling.

KW - Caveolin-1

KW - Clathrin

KW - Membrane docking

KW - Single-molecule fluorescence imaging

KW - SMAD4

UR - http://www.scopus.com/inward/record.url?scp=84888637943&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84888637943&partnerID=8YFLogxK

U2 - 10.1016/j.febslet.2013.10.041

DO - 10.1016/j.febslet.2013.10.041

M3 - Article

AN - SCOPUS:84888637943

SN - 0014-5793

VL - 587

SP - 3912

EP - 3920

JO - FEBS Letters

JF - FEBS Letters

IS - 24

ER -

Live-cell single-molecule imaging reveals clathrin and caveolin-1 dependent docking of SMAD4 at the cell membrane

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this