First published online January 11, 2006; 10.1104/pp.105.072678
Plant Physiology 140:726-733 (2006)
© 2006 American Society of Plant Biologists
OPEN ACCESS ARTICLE
CELL BIOLOGY AND SIGNAL TRANSDUCTION
Evidence for Functional Conservation, Sufficiency, and Proteolytic Processing of the CLAVATA3 CLE Domain1,[W],[OA]
Jun Ni and
Steven E. Clark*
Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109–1048
Arabidopsis (Arabidopsis thaliana) CLAVATA3 (CLV3) is hypothesized to act as a ligand for the CLV1 receptor kinase in the regulation of stem cell specification at shoot and flower meristems. CLV3 is a secreted protein, with an amino-terminal signal sequence and a conserved C-terminal domain of 15 amino acids, termed the CLE (CLV3/ESR-related) domain, based on its similarity to a largely unstudied protein family broadly present in land plants. We have tested the function of 13 Arabidopsis CLEs in vivo and found a significant variability in the ability of CLEs to replace CLV3, ranging from complete to no complementation. The best rescuing CLE depends on CLV1 for function, while other CLEs act independently of CLV1. Domain-swap experiments indicate that differences in function can be traced to the CLE domain within these proteins. Indeed, when the CLE domain of CLV3 is placed downstream of an unrelated signal sequence, it is capable of fully replacing CLV3 function. Finally, we have detected proteolytic activity in extracts from cauliflower (Brassica oleracea) that process both CLV3 and CLE1 at their C termini. For CLV3, processing appears to occur at the absolutely conserved arginine-70 found at the beginning of the CLE domain. We propose that CLV3 and other CLEs are C-terminally processed to generate an active CLE peptide.
1 This work was supported by the National Institutes of Health (grant no. 1R01GM62962–01A1 to S.E.C.).
The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Steven E. Clark (clarks{at}umich.edu).
[W] The online version of this article contains Web-only data.
[OA] Open Access articles can be viewed online without a subscription.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.072678.
* Corresponding author; e-mail clarks{at}umich.edu; fax 734–647–0884.
Received October 7, 2005;
returned for revision November 4, 2005;
accepted November 4, 2005.
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