Target of rapamycin regulates development and ribosomal RNA expression through kinase domain in Arabidopsis.
Target of rapamycin (TOR) is a central regulator of cell growth, cell death, nutrition, starvation, hormone, and stress responses in diverse eukaryotes. However, very little is known about TOR signaling and the associated functional domains in plants. We have taken a genetic approach to dissect TOR functions in Arabidopsis (Arabidopsis thaliana) and report here that the kinase domain is essential for the role of TOR in embryogenesis and 45S rRNA expression. Twelve new T-DNA insertion mutants, spanning 14.2 kb of TOR-encoding genomic region, have been characterized. Nine of these share expression of defective kinase domain and embryo arrest at 16 to 32 cell stage. However, three T-DNA insertion lines affecting FATC domain displayed normal embryo development, indicating that FATC domain was dispensable in Arabidopsis. Genetic complementation showed that the TOR kinase domain alone in tor-10/tor-10 mutant background can rescue early embryo lethality and restore normal development. Overexpression of full-length TOR or kinase domain in Arabidopsis displayed developmental abnormalities in meristem, leaf, root, stem, flowering time, and senescence. We further show that TOR, especially the kinase domain, plays a role in ribosome biogenesis by activating 45S rRNA production. Of the six putative nuclear localization sequences in the kinase domain, nuclear localization sequence 6 was identified to confer TOR nuclear targeting in transient expression assays. Chromatin immunoprecipitation studies revealed that the HEAT repeat domain binds to 45S rRNA promoter and the 5' external transcribed spacer elements motif. Together, these results show that TOR controls the embryogenesis, postembryonic development, and 45S rRNA production through its kinase domain in Arabidopsis.
Plant Biotechnology Institute, National Research Council of Canada, Saskatoon, Saskatchewan, Canada S7N 0W9.
Maozhi
M
Ren
Shuqing
S
Qiu
Prakash
P
Venglat
Daoquan
D
Xiang
Li
L
Feng
Gopalan
G
Selvaraj
Raju
R
Datla
D017360:Arabidopsis / Q000196:embryology / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D029681:Arabidopsis Proteins; D020029:Base Pairing / Q000235:genetics; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics; D018744:DNA, Plant / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant / Q000235:genetics; D016350:Leucine Zippers / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000235:genetics; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D019869:Phosphatidylinositol 3-Kinases; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012335:RNA, Ribosomal / Q000235:genetics* / Q000378:metabolism; D012639:Seeds / Q000201:enzymology / Q000254:growth & development; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic
D029681:Arabidopsis Proteins; D004269:DNA, Bacterial; D021901:DNA, Intergenic; D018744:DNA, Plant; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D012335:RNA, Ribosomal; C088179:RNA, ribosomal, 45S; C036483:T-DNA; C512171:TOR protein, Arabidopsis
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