Rap1 GTPase Regulation of Adherens Junction Positioning and Cell Adhesion

American Association for the Advancement of Science (AAAS) - Tập 295 Số 5558 - Trang 1285-1288 - 2002
Andrea L. Knox1, Nicholas H. Brown1
1Wellcome/CRC Institute and Department of Anatomy, University of Cambridge, Cambridge CB2 1QR, UK.

Tóm tắt

Cell-cell junctions are distributed evenly around the lateral circumference of cells within an epithelium. We find that the even distribution of adherens junctions is an active process that requires the small guanosine triphosphatase Rap1. Cells mutant for Rap1 condensed their adherens junctions to one side of the cell. This disrupted normal epithelial cell behavior, and mutant cell clones dispersed into the surrounding wild-type tissue. Rap1 is enriched at adherens junctions, particularly between newly divided sister cells where it may reseal the adherens junction ring. The regulation of adherens junction positioning could play a role in cell mobility and cell division.

Từ khóa


Tài liệu tham khảo

Kusumi A., Suzuki K., Koyasako K., Curr. Opin. Cell Biol. 11, 582 (1999).

Muller H. A., Dev. Dyn. 218, 52 (2000).

Bokoch G. M., Biochem. J. 289, 17 (1993).

Asha H., deRuiter N. D., Wang M. G., Hariharan I. K., EMBO J. 18, 605 (1999).

Garcia-Bellido A., Ripoll P., Morata G., Nature 245, 251 (1973).

Wild-type clones were generated by heat shocking y w P{ry +t7.2 hsFLP}22; P{w hs FRT}2A/P{w + nls-GFP} P{w hs FRT}2A flies and Rap1 clones by heat shocking y w P{ry +t7.2 hsFLP}22; Rap1 P[5709] P{w hs FRT}2A/ P{w + nls-GFP} P{w hs FRT}2A or y w P{ry +t7.2 hsFLP}22; Rap1 B1 P{w hs FRT}2A/ P{w + nls-GFP} P{w hs FRT}2A flies (25 26). Identical results were obtained with two amorphic alleles of Rap1 Rap1 B1 (4) and a P-element inserted into the coding region Rap1 P[5709] (23). For actin staining of pupal wings 30 to 36 hours after puparium formation (APF) (27) we used rhodamine-conjugated phalloidin (Molecular Probes). For antibody stains (27) we used DCAD2 anti–DE-cadherin (28) anti-cno (14) anti-ZO-1 (29) and anti-Dlg (30). All wings were fixed and stained at 30 to 36 hours APF except for wings stained for canoe which were aged 24 hours APF. Images were visualized with a MRC1024 confocal microscope (Bio-Rad).

Diaz-Benjumea F. J., Hafen E., Development 120, 569 (1994).

Strutt D. I., Weber U., Mlodzik M., Nature 387, 292 (1997).

Supplemental data are available on Science Online at www.sciencemag.org/cgi/content/full/295/5558/1285/DC1.

A. L. Knox N.H. Brown unpublished observations.

The size of groups of dispersed Rap1 mutant cells was counted in 23 clones from 21 different wings aged between 24 and 36 hours APF. The number of dispersed groups of a given size (and fraction of the total 244 groups) was as follows: 13 (5%) single cells; 106 (43%) cell pairs; 20 (8%) 3-cell groups; 56 (23%) 4-cell groups; 8 (3%) 5-cell groups; 16 (7%) 6-cell groups; and 25 (10%) groups of 7 or more cells.

T. Linnemann et al. J. Biol. Chem. 274 13556 (1999).

B. Boettner L. VanAelst U. Gaul personal communication

Takahashi K., Matsuo T., Katsube T., Ueda R., Yamamoto D., Mech. Dev. 78, 97 (1998).

Itoh M., Nagafuchi A., Moroi S., Tsukita S., J. Cell Biol. 138, 181 (1997).

Wei X., Ellis H. M., Mech. Dev. 100, 217 (2001).

Stevenson B. R., Siliciano J. D., Mooseker M. S., Goodenough D. A., J. Cell Biol. 103, 755 (1986).

Steinberg M. S., Takeichi M., Proc. Natl. Acad. Sci. U.S.A. 91, 206 (1994).

Godt D., Tepass U., Nature 395, 387 (1998).

Gonzalez-Reyes A., St Johnston D., Development 125, 3635 (1998).

D. Fristrom J. W. Fristrom in The Development of Drosophila melanogaster M. Bate A. Martinez Arias Eds. (Cold Spring Harbor Laboratory Press Cold Spring Harbor NY 1993) vol. 2 pp. 843–896.

Rap1 mutant wing clones expressing α-catenin–GFP (31) were generated by heat shocking y w P{ry +t7.2 hsFLP}22; P{UAS-DαC-GFP#3} {arm-GAL4}/+; Rap1 P[5709] P{w hs FRT}2A/ P{w + nls-GFP} P{w hs FRT}2A flies. 24h APF pupae were dissected from the pupal case and directly mounted in Vectashield (Vector Labs Burlingame CA) then clones were imaged with a MRC1024 confocal microscope (Bio-Rad). Rap1 was tagged with GFP by generating a genomic rescue construct with GP inserted at the NH 2 -terminus of Rap1 (23). Imaginal discs and 2-hour APF wings expressing GFP-Rap1 or α-catenin–GFP were fixed in 4% formaldehyde and mounted in Vectashield. Images were collected with a Radiance 2000 confocal microscope (Bio-Rad).

A. L. Knox N. H. Brown in preparation.

Woods D. F., Wu J. W., Bryant P. J., Dev. Genet. 20, 111 (1997).

10.1093/genetics/144.4.1673

10.1016/S1097-2765(00)80419-0

Fristrom D., Wilcox M., Fristrom J., Development 117, 509 (1993).

Oda H., Uemura T., Harada Y., Iwai Y., Takeichi M., Dev. Biol. 165, 716 (1994).

M. Takahisa et al. Genes Dev. 10 1783 (1996).

10.1016/0092-8674(81)90009-X

Oda H., Tsukita S., Dev. Genes Evol. 209, 218 (1999).

We thank P. Bryant R. Fehon I. Hariharan R. Howes H. Oda S. Luschnig M. Takahisa M. Takeichi and D. Yamamoto for providing antibodies and fly stocks and S. Bray B. Harris D. St Johnston and R. White for critically reading the manuscript. We thank J. Overton for technical assistance and the members of our lab for valuable advice. Supported by a Wellcome Trust Senior Fellowship (N.H.B.) and grants from the Association of Commonwealth Universities the New Zealand Federation of University Women and Trinity College (A.L.K.).

Thông tin
Thông tin xuất bản

American Association for the Advancement of Science (AAAS)

Tập 295 Số 5558

1285-1288

Thông tin tác giả