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Published on February 9, 2007; 10.1104/pp.106.092296

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Received October 30, 2006
Accepted January 31, 2007

An Early Nodulin-like Protein Accumulates in the Sieve Element Plasma Membrane of Arabidopsis

Junaid A. Khan , Qi Wang , Richard D. Sjölund , Alexander Schulz , and Gary A. Thompson *

Department of Applied Science, University of Arkansas at Little Rock, 2801 S. University Ave. Little Rock, Arkansas 72204-1099 USA; Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242 USA; Department of Plant Biology, Faculty of Life Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark

* Corresponding author; email: gathompson{at}ualr.edu.

Membrane proteins within the sieve element-companion cell complex have essential roles in the physiological functioning of the phloem. The monoclonal antibody line RS6, selected from hybridomas raised against sieve elements isolated from Streptanthus tortuosus (California shield leaf, Brassicaceae) tissue cultures, recognizes an antigen in the Arabidopsis thaliana ecotype Columbia that is associated specifically with the plasma membrane of sieve elements, but not companion cells, and accumulates at the earliest stages of sieve element differentiation. The identity of the RS6 antigen was revealed by RT-PCR of Arabidopsis leaf RNA using degenerate primers to be an early nodulin (ENOD)-like protein that is encoded by the expressed gene At3g20570. Arabidopsis ENOD-like proteins are encoded by a multigene family composed of several types of structurally related phytocyanins that have a similar overall domain structure of an amino-terminal signal peptide, plastocyanin-like copper binding domain, proline/serine-rich domain and carboxy-terminal hydrophobic domain. The amino- and carboxy-terminal domains of the 21.5 kDa SE-ENOD are post-translationally cleaved from the precursor protein resulting in a mature peptide of ~15 kDa that is attached to the sieve element plasma membrane via a carboxy-terminal glycosylphosphatidylinositol (GPI) membrane anchor. Many of the Arabidopsis ENOD-like proteins accumulate in gametophytic tissues, whereas in both floral and vegetative tissues, the SE-ENOD is expressed only within the phloem. Members of the ENOD subfamily of the cupredoxin superfamily do not appear to bind copper and have unknown functions. Phenotypic analysis of homozygous T-DNA insertion mutants for the gene At3g20570 shows minimal alteration in vegetative growth, but a significant reduction in the overall reproductive potential.




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