Multi-drug resistance protein 4 (MRP4)-mediated regulation of fibroblast cell migration reflects a dichotomous role of intracellular cyclic nucleotides

Chandrima Sinha, Aixia Ren, Kavisha Arora, Chang Suk Moon, Sunitha Yarlagadda, Weiqiang Zhang, Satish B. Cheepala, John D. Schuetz, Anjaparavanda P. Naren

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

It has long been known that cyclic nucleotides and cyclic nucle-otide-dependent signaling molecules control cell migration. However, the concept that it is not just the absence or presence of cyclic nucleotides, but a highly coordinated balance between these molecules that regulates cell migration, is new and revolutionary. In this study, we used multidrug resistance protein 4 (MRP4)-ex-pressing cell lines and MRP4 knock-out mice as model systems and wound healing assays as the experimental system to explore this unique and emerging concept. MRP4, a member of a large family of ATP binding cassette transporter proteins, localizes to the plasma membrane and functions as a nucleotide efflux transporter and thus plays a role in the regulation of intracellular cyclic nucleotide levels. Here, we demonstrate that mouse embryonic fibroblasts (MEFs) isolated from Mrp4-/- mice have higher intracellular cyclic nucleotide levels and migrate faster compared with MEFs from Mrp4+/+ mice. Using FRET-based cAMP and cGMP sensors, we show that inhibition of MRP4 with MK571 increases both cAMP and cGMP levels, which results in increased migration. In contrast to these moderate increases in cAMP and cGMP levels seen in the absence of MRP4, a robust increase in cAMP levels was observed following treatment with forskolin and isobutylmethylx-anthine, which decreases fibroblast migration. In response to externally added cell-permeant cyclic nucleotides (cpt-cAMP and cpt-cGMP), MEF migration appears to be biphasic. Altogether, our studies provide the first experimental evidence supporting the novel concept that balance between cyclic nucleotides is critical for cell migration.

Original languageEnglish (US)
Pages (from-to)3786-3794
Number of pages9
JournalJournal of Biological Chemistry
Volume288
Issue number6
DOIs
StatePublished - Feb 8 2013

Fingerprint

Cyclic Nucleotides
Multiple Drug Resistance
Fibroblasts
Cell Movement
Cells
Pharmaceutical Preparations
Proteins
P-Glycoproteins
Molecules
ATP-Binding Cassette Transporters
Colforsin
Cell membranes
Knockout Mice
Wound Healing
Assays
Nucleotides
Cell Membrane
Cell Line
Sensors

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Multi-drug resistance protein 4 (MRP4)-mediated regulation of fibroblast cell migration reflects a dichotomous role of intracellular cyclic nucleotides. / Sinha, Chandrima; Ren, Aixia; Arora, Kavisha; Moon, Chang Suk; Yarlagadda, Sunitha; Zhang, Weiqiang; Cheepala, Satish B.; Schuetz, John D.; Naren, Anjaparavanda P.

In: Journal of Biological Chemistry, Vol. 288, No. 6, 08.02.2013, p. 3786-3794.

Research output: Contribution to journalArticle

Sinha, Chandrima ; Ren, Aixia ; Arora, Kavisha ; Moon, Chang Suk ; Yarlagadda, Sunitha ; Zhang, Weiqiang ; Cheepala, Satish B. ; Schuetz, John D. ; Naren, Anjaparavanda P. / Multi-drug resistance protein 4 (MRP4)-mediated regulation of fibroblast cell migration reflects a dichotomous role of intracellular cyclic nucleotides. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 6. pp. 3786-3794.
@article{902115daf8ca40189a8b5989eb910757,
title = "Multi-drug resistance protein 4 (MRP4)-mediated regulation of fibroblast cell migration reflects a dichotomous role of intracellular cyclic nucleotides",
abstract = "It has long been known that cyclic nucleotides and cyclic nucle-otide-dependent signaling molecules control cell migration. However, the concept that it is not just the absence or presence of cyclic nucleotides, but a highly coordinated balance between these molecules that regulates cell migration, is new and revolutionary. In this study, we used multidrug resistance protein 4 (MRP4)-ex-pressing cell lines and MRP4 knock-out mice as model systems and wound healing assays as the experimental system to explore this unique and emerging concept. MRP4, a member of a large family of ATP binding cassette transporter proteins, localizes to the plasma membrane and functions as a nucleotide efflux transporter and thus plays a role in the regulation of intracellular cyclic nucleotide levels. Here, we demonstrate that mouse embryonic fibroblasts (MEFs) isolated from Mrp4-/- mice have higher intracellular cyclic nucleotide levels and migrate faster compared with MEFs from Mrp4+/+ mice. Using FRET-based cAMP and cGMP sensors, we show that inhibition of MRP4 with MK571 increases both cAMP and cGMP levels, which results in increased migration. In contrast to these moderate increases in cAMP and cGMP levels seen in the absence of MRP4, a robust increase in cAMP levels was observed following treatment with forskolin and isobutylmethylx-anthine, which decreases fibroblast migration. In response to externally added cell-permeant cyclic nucleotides (cpt-cAMP and cpt-cGMP), MEF migration appears to be biphasic. Altogether, our studies provide the first experimental evidence supporting the novel concept that balance between cyclic nucleotides is critical for cell migration.",
author = "Chandrima Sinha and Aixia Ren and Kavisha Arora and Moon, {Chang Suk} and Sunitha Yarlagadda and Weiqiang Zhang and Cheepala, {Satish B.} and Schuetz, {John D.} and Naren, {Anjaparavanda P.}",
year = "2013",
month = "2",
day = "8",
doi = "10.1074/jbc.M112.435925",
language = "English (US)",
volume = "288",
pages = "3786--3794",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "6",

}

TY - JOUR

T1 - Multi-drug resistance protein 4 (MRP4)-mediated regulation of fibroblast cell migration reflects a dichotomous role of intracellular cyclic nucleotides

AU - Sinha, Chandrima

AU - Ren, Aixia

AU - Arora, Kavisha

AU - Moon, Chang Suk

AU - Yarlagadda, Sunitha

AU - Zhang, Weiqiang

AU - Cheepala, Satish B.

AU - Schuetz, John D.

AU - Naren, Anjaparavanda P.

PY - 2013/2/8

Y1 - 2013/2/8

N2 - It has long been known that cyclic nucleotides and cyclic nucle-otide-dependent signaling molecules control cell migration. However, the concept that it is not just the absence or presence of cyclic nucleotides, but a highly coordinated balance between these molecules that regulates cell migration, is new and revolutionary. In this study, we used multidrug resistance protein 4 (MRP4)-ex-pressing cell lines and MRP4 knock-out mice as model systems and wound healing assays as the experimental system to explore this unique and emerging concept. MRP4, a member of a large family of ATP binding cassette transporter proteins, localizes to the plasma membrane and functions as a nucleotide efflux transporter and thus plays a role in the regulation of intracellular cyclic nucleotide levels. Here, we demonstrate that mouse embryonic fibroblasts (MEFs) isolated from Mrp4-/- mice have higher intracellular cyclic nucleotide levels and migrate faster compared with MEFs from Mrp4+/+ mice. Using FRET-based cAMP and cGMP sensors, we show that inhibition of MRP4 with MK571 increases both cAMP and cGMP levels, which results in increased migration. In contrast to these moderate increases in cAMP and cGMP levels seen in the absence of MRP4, a robust increase in cAMP levels was observed following treatment with forskolin and isobutylmethylx-anthine, which decreases fibroblast migration. In response to externally added cell-permeant cyclic nucleotides (cpt-cAMP and cpt-cGMP), MEF migration appears to be biphasic. Altogether, our studies provide the first experimental evidence supporting the novel concept that balance between cyclic nucleotides is critical for cell migration.

AB - It has long been known that cyclic nucleotides and cyclic nucle-otide-dependent signaling molecules control cell migration. However, the concept that it is not just the absence or presence of cyclic nucleotides, but a highly coordinated balance between these molecules that regulates cell migration, is new and revolutionary. In this study, we used multidrug resistance protein 4 (MRP4)-ex-pressing cell lines and MRP4 knock-out mice as model systems and wound healing assays as the experimental system to explore this unique and emerging concept. MRP4, a member of a large family of ATP binding cassette transporter proteins, localizes to the plasma membrane and functions as a nucleotide efflux transporter and thus plays a role in the regulation of intracellular cyclic nucleotide levels. Here, we demonstrate that mouse embryonic fibroblasts (MEFs) isolated from Mrp4-/- mice have higher intracellular cyclic nucleotide levels and migrate faster compared with MEFs from Mrp4+/+ mice. Using FRET-based cAMP and cGMP sensors, we show that inhibition of MRP4 with MK571 increases both cAMP and cGMP levels, which results in increased migration. In contrast to these moderate increases in cAMP and cGMP levels seen in the absence of MRP4, a robust increase in cAMP levels was observed following treatment with forskolin and isobutylmethylx-anthine, which decreases fibroblast migration. In response to externally added cell-permeant cyclic nucleotides (cpt-cAMP and cpt-cGMP), MEF migration appears to be biphasic. Altogether, our studies provide the first experimental evidence supporting the novel concept that balance between cyclic nucleotides is critical for cell migration.

UR - http://www.scopus.com/inward/record.url?scp=84873660811&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84873660811&partnerID=8YFLogxK

U2 - 10.1074/jbc.M112.435925

DO - 10.1074/jbc.M112.435925

M3 - Article

VL - 288

SP - 3786

EP - 3794

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 6

ER -