Assembly of adherens junctions is required for sphingosine 1-phosphate-induced matriptase accumulation and activation at mammary epithelial cell-cell contacts.
Sphingosine 1-phosphate (S1P), a bioactive phospholipid, simultaneously induces actin cytoskeletal rearrangements and activation of matriptase, a membrane-associated serine protease in human mammary epithelial cells. In this study, we used a monoclonal antibody selective for activated, two-chain matriptase to examine the functional relationship between these two S1P-induced events. Ten minutes after exposure of 184 A1N4 mammary epithelial cells to S1P, matriptase was observed to accumulate at cell-cell contacts. Activated matriptase first began to appear as small spots at cell-cell contacts, and then its deposits elongated along cell-cell contacts. Concomitantly, S1P induced assembly of adherens junctions and subcortical actin belts. Matriptase localization was observed to be coincident with markers of adherens junctions at cell-cell contacts but likely not to be incorporated into the tightly bound adhesion plaque. Disruption of subcortical actin belt formation and prevention of adherens junction assembly led to prevention of accumulation and activation of the protease at cell-cell contacts. These data suggest that S1P-induced accumulation and activation of matriptase depend on the S1P-induced adherens junction assembly. Although MAb M32, directed against one of the low-density lipoprotein receptor class A domains of matriptase, blocked S1P-induced activation of the enzyme, the antibody had no effect on S1P-induced actin cytoskeletal rearrangement. Together, these data indicate that actin cytoskeletal rearrangement is necessary but not sufficient for S1P-induced activation of matriptase at cell-cell contacts. The coupling of matriptase activation to adherens junction assembly and actin cytoskeletal rearrangement may serve to ensure tight control of matriptase activity, restricted to cell-cell junctions of mammary epithelial cells.
Hung
Ruei-Jiun
RJ
Lombardi Cancer Center, Georgetown Univ. Medical Center 3970 Reservoir Rd. NW, Washington, DC 20057-1412, USA.
Hsu
Ia-Wen J
IaW
Dreiling
Jennifer L
JL
Lee
Mon-Juan
MJ
Williams
Cicely A
CA
Oberst
Michael D
MD
Dickson
Robert B
RB
Lin
Chen-Yong
CY
D000199:Actins / Q000378:metabolism; D022005:Adherens Junctions / Q000502:physiology*; D001940:Breast / Q000201:enzymology / Q000502:physiology*; D002450:Cell Communication / Q000502:physiology*; D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism; D004789:Enzyme Activation / Q000502:physiology; D004847:Epithelial Cells / Q000201:enzymology / Q000502:physiology; D005260:Female; D006801:Humans; D007700:Kinetics; D008246:Lysophospholipids / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000502:physiology; D012697:Serine Endopeptidases; D013110:Sphingosine / Q000031:analogs & derivatives / Q000502:physiology*
D000199:Actins; D008246:Lysophospholipids; C060506:sphingosine 1-phosphate; D012697:Serine Endopeptidases; C485595:matriptase; D013110:Sphingosine