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J. Biol. Chem., Vol. 277, Issue 12, 9840-9852, March 22, 2002
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From the Department of Biology and Program in Cell and Molecular
Biology, Colorado State University, Fort Collins, Colorado 80523
Analysis of the recently completed
Arabidopsis genome sequence indicates that ~31% of the
predicted genes could not be assigned to functional categories,
as they do not show any sequence similarity with proteins of known
function from other organisms. Calmodulin (CaM), a ubiquitous and
multifunctional Ca2+ sensor, interacts with a wide variety
of cellular proteins and modulates their activity/function in
regulating diverse cellular processes. However, the primary
amino acid sequence of the CaM-binding domain in different CaM-binding
proteins (CBPs) is not conserved. One way to identify most of the CBPs
in the Arabidopsis genome is by protein-protein
interaction-based screening of expression libraries with CaM. Here,
using a mixture of radiolabeled CaM isoforms from
Arabidopsis, we screened several expression libraries prepared from flower meristem, seedlings, or tissues treated with hormones, an elicitor, or a pathogen. Sequence analysis of 77 positive
clones that interact with CaM in a
Ca2+-dependent manner revealed 20 CBPs,
including 14 previously unknown CBPs. In addition, by searching the
Arabidopsis genome sequence with the newly identified and
known plant or animal CBPs, we identified a total of 27 CBPs. Among
these, 16 CBPs are represented by families with 2-20 members in each
family. Gene expression analysis revealed that CBPs and CBP paralogs
are expressed differentially. Our data suggest that
Arabidopsis has a large number of CBPs including several
plant-specific ones. Although CaM is highly conserved between plants
and animals, only a few CBPs are common to both plants and animals.
Analysis of Arabidopsis CBPs revealed the presence of a
variety of interesting domains. Our analyses identified several
hypothetical proteins in the Arabidopsis genome as CaM targets, suggesting their involvement in Ca2+-mediated
signaling networks.
Genes Encoding Calmodulin-binding Proteins in the
Arabidopsis Genome*,
,, and
*
This work was supported in part by grants from the National
Science Foundation and the National Aeronautics and Space
Administration (to A. S. N. R.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
The on-line version of this article (available at
http://www.jbc.org) contains Table IV.
These authors contributed equally to this work.
§
To whom correspondence should be addressed. Tel.: 970-491-5773;
Fax: 970-491-0649; E-mail: reddy@lamar.colostate.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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