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

First published online 21 March 2007
doi: 10.1242/dev.001016

This Article Summary Full Text Full Text (PDF) Supplementary Material 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 HighWire Citing Articles via Google Scholar Google Scholar Articles by Chandler, J. W. Articles by Werr, W. Search for Related Content PubMed PubMed Citation Articles by Chandler, J. W. Articles by Werr, W. Social Bookmarking                         What's this?

The AP2 transcription factors DORNRÖSCHEN and DORNRÖSCHEN-LIKE redundantly control Arabidopsis embryo patterning via interaction with PHAVOLUTA

Institute of Developmental Biology, University of Cologne, Gyrhofstrasse 17, D-50923, Cologne, Germany.

View larger version (68K): [in this window] [in a new window]   Fig. 1. drn mutant and drn drnl double-mutant phenotypes. Wildtype embryos at early (A) and late (B) globular stages. Abnormal hypophyseal cell divisions in drn-1 mutant embryos at early (C) and late (D) globular stages. (E) Multiple cell files in the suspensor of a drn1 mutant; the embryo proper is towards the top. (F) drn-1 mutants lacking basal embryo organisation. (G-K) Pleiotropy of cotyledon phenotypes for the drn-1 mutant showing mono-(G) and polycotyledony (I), cotyledon fusion (H) and cup-shaped cotyledons (J). The SAM is active despite complete cotyledon fusion (K). (L,M) mp-like phenotype of drn (L) and drn-1 drnl-1 (M) mutants. (N,O) Altered phyllotaxis of first leaves in monocotyledonous (N) or polycotyledonous (O) drn-1 mutants. (P-R) Cleared cotyledons of wild type (P) and a monocotyledonous drn-1 mutant showing single (Q) or fused cotyledon (R) ontogeny. (S,T) drn-1 drnl-1 double-mutant embryos showing asymmetric cell divisions at early (S) or late (T) globular stages. (U,V) pin-like hypocotyl of drn-1 drnl-2 double-mutant plant lacking cotyledons (U) and directly producing leaves as evidenced by trichome formation (V).

View larger version (79K): [in this window] [in a new window]   Fig. 2. RNA in situ hybridisations and alteration of DR5 and PIN1 expression in drn mutants. (A-D) DRN expression on median longitudinal sections of wild-type embryos in early globular (A), late globular (B), early heart (C) and torpedo (D) stages. (E-H) DRNL expression in wild-type embryos in early globular (E), late globular (F), early heart (G) and late heart (H) stages. (I-K) DR5::GFP expression in globular wild-type (I), globular drn-1 mutant (J) and drn-1 transition-stage (K) embryos. The arrow in J marks the hypophysis region. Note supernumery cell files in the suspensor (K). (L,M) DR5::GFP expression in wild-type (L) and drn-1 (M) heart-stage embryos. Note the normal and mutant cotyledons in M. (N-Q) PIN1 expression in wild-type (N) and drn (O) 32-cell-stage embryos, and in wild-type (P) and drn (Q) globular embryos. Arrows show basal PIN1 polarity (N,P) and abnormal lateral PIN1 expression (O,Q). (R-U) RNA in situ hybridisation for PHV on median longitudinal sections of wild-type embryos at early globular (R), late globular (S), heart (T) or late heart (U) stage.

View larger version (24K): [in this window] [in a new window]   Fig. 3. CoIP and BiFC analysis for the interaction between DRN or DRNL and PHV. (A,B) Full-length PHV protein can precipitate fulllength DRN (A) or DRNL (B). Protein mixtures were precipitated via PHV-HA. The grid summarises the IP and Co-IP lanes and the antibodies used. Asterisks mark the co-eluted IgG light chain. (C-E) Cellular fluorescence of GFP-DRN (C), GFP-DRNL (D) or GFP-PHVs (E) in leek epidermal cells. (F,G) Bimolecular fluorescence complementation showing YFP expression observed with a GFP filter in leek epidermal cells following co-bombardment of full-length DRN and PHVs proteins (F) or full-length DRNL and PHVs proteins (G) fused to complementary YFP subdomains.

View larger version (34K): [in this window] [in a new window]   Fig. 4. The interaction between DRN and class III HD-ZIP family proteins involves the AP2 and PAS-like domains, respectively. (A)A comparison of the C-terminal regions of the Arabidopsis class III HD-ZIP proteins. Accession numbers for the proteins are as follows: CNA, AAW88440; AtHB8, CAD29660; REV AAF42938; PHV CAD29544; PHB, NP_181018. Homology is compared over the region from PHV amino acid 654 to the C-terminus. The MEKHLA domain, homology to the PAS domain from http://smart.embl-heidelberg.de and the -helices within the PHV PAS domain are marked. (B) Full-length DRN is precipitated by C-terminal regions of all class III HD-ZIP members, but not by DRNL. HA-tagged class III HD-ZIP proteins or DRNL were used for precipitation of DRN. The grid shows IP and Co-IP lanes and which antibodies were used. The position of the DRN protein is marked with an arrowhead. Asterisks represent the IgG heavy or light chains. The control lanes show IPs in the absence of either or both in vitro transcribed/translated products, confirming the identity of the lower IgG band. (C) Full-length PHV cannot co-precipitate full-length STM-GFP. The first and third lanes contain in vitro translated proteins before precipitation. PHV-HA was used for precipitation. (D) Full-length DRN can co-precipitate the 71 amino acid PHVPAS domain (first and second lanes) and, reciprocally, PHVPAS can co-precipitate full-length DRNL (third and fourth lanes). (E) An alignment of the AP2 domains of DRN and DRNL. Identical amino acids are shaded and the position of the unique cysteine and serine residues within the -helix are in bold. (F) The DRN AP2 domain (IP) can precipitate PHVs (Co-IP) (first and third lanes) and the DRNL AP2 domain can precipitate PHVs (second and fourth lanes). (G) 3D crystal structure of the AP2 domain of the Arabidopsis ERF1 protein (Allen et al., 1998; Marchler-Bauer et al., 2005) binding a DNA helix shown on the right, viewed from above into the helix. Cysteine/serine residues of the DRN/DRNL proteins are highlighted in yellow.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

Twitter