Ubiquitin C-terminal hydrolases 1 and 2 affect shoot architecture in Arabidopsis.
Ubiquitin C-terminal hydrolases (UCHs) are a subset of de-ubiquitinating proteases that release covalently linked ubiquitin (Ub), and as such play essential roles in recycling Ub and reversing the action of Ub conjugation. We show here that two related Arabidopsis UCHs, UCH1, and UCH2, are important for shoot development. The UCH1 and 2 genes are ubiquitously expressed, with the corresponding proteins present in both the cytoplasm and nucleus. Unlike their animal and fungal counterparts, we found no evidence that the Arabidopsis UCH1 and 2 proteins stably associate with the 26S proteasome. Altering the levels of UCH1 and 2 has substantial effects on Arabidopsis shoot development, especially with respect to inflorescence architecture, with over-expression and double mutants enhancing and suppressing the outgrowth of cauline branches, respectively. Neither UCH1-over-expressing nor uch1-1 uch2-1 plants have detectably altered sensitivity to cytokinins or auxins individually, but exhibit an altered sensitivity to the ratio of the two hormones. UCH1-over-expressing plants show dramatically enhanced phenotypes when combined with auxin-insensitive mutants axr1-3 and axr2-1, suggesting that one or more aspects of auxin signaling are affected by this enzyme pair. Previous studies revealed that the ubiquitination and degradation of the AUX/IAA family of repressors is a key step in auxin signaling. Here, we show that turnover of a reporter fused to a representative AUX/IAA protein AXR3 is faster in the uch1-1 uch2-1 double mutant but slower in the UCH1 over-expression backgrounds. Taken together, our results indicate that de-ubiquitination helps to modify plant shoot architecture, possibly via its ability to directly or indirectly protect upstream target proteins involved in auxin/cytokinin signaling from Ub-mediated degradation.
Yang
Peizhen
P
Department of Genetics, University of Wisconsin-Madison, Madison, WI 53706-1574, USA.
Smalle
Jan
J
Lee
Sangsook
S
Yan
Ning
N
Emborg
Thomas J
TJ
Vierstra
Richard D
RD
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000254:growth & development / Q000502:physiology; D029681:Arabidopsis Proteins / Q000378:metabolism; D017796:Caulimovirus; D003583:Cytokinins / Q000378:metabolism; D015870:Gene Expression; D007210:Indoleacetic Acids / Q000378:metabolism*; D008969:Molecular Sequence Data; D005810:Multigene Family; D009154:Mutation; D010641:Phenotype; D018520:Plant Shoots / Q000254:growth & development*; D011401:Promoter Regions, Genetic; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D025801:Ubiquitin / Q000378:metabolism; D043222:Ubiquitin Thiolesterase / Q000378:metabolism*; D014452:Ubiquitins / Q000378:metabolism
D029681:Arabidopsis Proteins; D003583:Cytokinins; D007210:Indoleacetic Acids; C513170:RUB1 protein, Arabidopsis; D025801:Ubiquitin; D014452:Ubiquitins; D043222:Ubiquitin Thiolesterase; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease