QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online 6 July 2005
doi: 10.1242/dev.01930

This Article Summary Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Wang, J. Articles by Savage-Dunn, C. Search for Related Content PubMed PubMed Citation Articles by Wang, J. Articles by Savage-Dunn, C. Social Bookmarking                         What's this?

C-terminal mutants of C. elegans Smads reveal tissue-specific requirements for protein activation by TGF-ß signaling

¹ Department of Biology, Queens College, and the Graduate School and University Center, the City University of New York, 65-30 Kissena Boulevard, Flushing, NY 11367, USA
² Department of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA

View larger version (56K): [in a new window]   Fig. 1. Expression patterns of Smad constructs in vivo. (A) Localization of SMA-3 mutant forms. The SMA-3 AMA protein accumulates in the nucleus, similar to the accumulation of wild-type SMA-3. The other dominant-negative SMA-3 mutants have a similar localization pattern (data not shown). (B) The expression pattern of a sma-2::gfp transcriptional fusion gene in (B1) the whole animal, (B2) pharynx, (B3) intestine and (B4) hypodermis. (C) A functional genomic clone of sma-2 excluding the two large introns.

View larger version (121K): [in a new window]   Fig. 2. The yeast two-hybrid test of the interaction between LIN-31 and SMA-3 or SMA-2. (A) An interaction can be detected between SMA-3 and LIN-31 by using a His selection plate (50 mM 3AT): (1) sma-3-AMA:lin-31, (2) sma-3-DME:lin-31, (3) sma-3:lin-31, (4) lin-31:vector, (5) sma-3-AMA:vector, (6) sma-3-DME:vector, (7) sma-3:vector and (8) vector:vector. (B) There was no detectable interaction between SMA-2 and LIN-31 when using the His selection plate (25 mM 3AT). (1) sma-3:vector, (2) sma-3:lin-31, (3) lin-31:vector, (4) vector:vector, (5) sma-2:lin-31, (6) sma-2:vector, (7) sma-2-DID:lin-31 and (8) sma-2-DID:vector. (C) Both MH1 and MH2 domains of SMA-3 can interact with LIN-31, as detected using the His selection plate (25 mM 3AT): (1) sma-3:vector, (2) sma-3:lin-31, (3) lin-31:vector, (4) vector:vector, (5) sma-3 MH2:lin-31, (6) sma-3 MH2: vector, (7) sma-3 MH1:lin-31 and (8) sma-3 MH1:vector. Note, the sma-2 or sma-3 cDNA was inserted into pPDleu vector and lin-31 cDNA was inserted into pPC86 vector. The yeast strain used for all of the tests was Mav203.

View larger version (85K): [in a new window]   Fig. 3. The SMA-3 C-terminal mutants activate a negative-feedback loop. All of the sma-3 constructs were co-injected with sma-3::gfpC. (A) The fluorescence of sma-3::gfpC alone; (B) wild-type sma-3 activates the feedback loop, as does (C) sma-3 SMT, (D) sma-3 AMA and (E) sma-3 DME. All photographs were taken under the same conditions with the same exposure time.

View larger version (19K): [in a new window]   Fig. 4. Proposed model of the functional complexes of C. elegans Smads. The body size control requires a SMA-3:SMA-4:SMA-2 complex and the feedback loop needs a SMA-2:SMA-4:SMA-3 complex; subunits are shown in an A,B,C configuration, as defined by Chacko et al. (Chacko et al., 2004).

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

Twitter