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United States Department of Agriculture

Agricultural Research Service

Research Project: MOLECULAR DEVELOPMENTAL GENETICS OF POLLEN AND POLLEN-PISTIL INTERACTIONS IN CROP PLANTS

Location: Plant Gene Expression Center Albany_CA

Title: A distinct mechanism regulating a pollen-specific guanine nucleotide exchange factor for the small GTPase Rop in Arabidopsis thaliana

Authors
item Zhang, Y. - ARS UCB PLNT GENE EXP CT
item McCormick, Sheila

Submitted to: Proceedings of the National Academy of Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 28, 2007
Publication Date: November 20, 2007
Repository URL: http://www.pnas.org/content/104/47/18830.full.pdf+html
Citation: Zhang, Y., McCormick, S.M. 2007. A distinct mechanism regulating a pollen-specific guanine nucleotide exchange factor for the small GTPase Rop in Arabidopsis thaliana. Proceedings of the National Academy of Sciences. 104:18830-18835.

Interpretive Summary: We studied a protein called RopGEF that is important for pollen tube growth. If this protein is overexpressed in pollen tubes there is no obvious change, but if the back part of the protein is deleted, the pollen tubes no longer grow normally, but instead grow in all directions. These results indicate that the back part of the protein is important to inhibit the activity of ROPGEF spatially, so that the pollen tube grows only from the tip. We further showed that the back part of RopGEF interacts with a pollen receptor kinase.

Technical Abstract: Rop/Rac small GTPases are central to diverse developmental and cellular activities in plants, playing an especially important Role in polar growth of pollen tubes. Although it is established that a class of plant-specific RopGEFs promotes the activity of Rop/Rac through the catalytic PRONE (Plant-specific Rop nucleotide exchanger) domain, not much is known about how RopGEF function is controlled to allow a spatiotemporally regulated Rop activity. To understand such a process in pollen, we performed functional analysis with a pollen-specific RopGEF, AtRopGEF12. Overexpression of At-RopGEF12 had minimal phenotypic effects, whereas overexpression of a C-terminally truncated version disturbed tube growth, suggesting that the C terminus was inhibitory to GEF function. In contrast to non-pollen-expressed RopGEFs, pollen-expressed RopGEFs have conserved C termini. A phospho-mimicking mutation at an invariant serine within the C terminus of AtRopGEF12 resulted in loss of the C-terminal inhibition, suggesting that phosphorylation regulates GEF activity in vivo. The PRONE domain of AtRopGEF12 (PRONE12) was not sufficient to induce isotropic tube growth. We used mbSUS to show that AtRopGEF12 interacts with an Arabidopsis pollen receptor kinase AtPRK2a through its C terminus, and BiFC to show that they interact in pollen tubes. Coexpression of AtRopGEF12 and AtPRK2a caused isotropic growth reminiscent of that seen upon overexpression of a constitutively active (CA) Rop. Coexpression of AtPRK2a with an Nterminally truncated AtRopGEF12 did not induce isotropic growth, indicating a positive role for the N-terminal domain. Our results suggest a mechanism by which the noncatalytic domains of pollenspecific/enriched RopGEFs regulate PRONE function, leading to polarized pollen tube growth.

Last Modified: 10/1/2014
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