Large-scale analysis of the GRAS gene family in Arabidopsis thaliana.
GRAS proteins belong to a plant-specific transcription factor family. Currently, 33 GRAS members including a putative expressed pseudogene have been identified in the Arabidopsis genome. With a reverse genetic approach, we have constructed a "phenome-ready unimutant collection" of the GRAS genes in Arabidopsis thaliana. Of this collection, we focused on loss-of-function mutations in 23 novel GRAS members. Under standard conditions, homozygous mutants have no obvious morphological phenotypes compared with those of wild-type plants. Expression analysis of GRAS genes using quantitative real-time RT-PCR (qRT-PCR), microarray data mining, and promoter::GUS reporter fusions revealed their tissue-specific expression patterns. Our analysis of protein-protein interaction and subcellular localization of individual GRAS members indicated their roles as transcription regulators. In our yeast two-hybrid (Y2H) assay, we confirmed the protein-protein interaction between SHORT-ROOT (SHR) and SCARECROW (SCR). Furthermore, we identified a new SHR-interacting protein, SCARECROW-LIKE23 (SCL23), which is the most closely related to SCR. Our large-scale analysis provides a comprehensive evaluation on the Arabidopsis GRAS members, and also our phenome-ready unimutant collection will be a useful resource to better understand individual GRAS proteins that play diverse roles in plant growth and development.
Lee
Mi-Hyun
MH
Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701, Korea.
Kim
Bohye
B
Song
Sang-Kee
SK
Heo
Jung-Ok
JO
Yu
Nan-Ie
NI
Lee
Shin Ae
SA
Kim
Miran
M
Kim
Dong Gwan
DG
Sohn
Sung Oh
SO
Lim
Chae Eun
CE
Chang
Kwang Suk
KS
Lee
Myeong Min
MM
Lim
Jun
J
D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000145:classification / Q000235:genetics* / Q000502:physiology*; D002467:Cell Nucleus / Q000737:chemistry; D015870:Gene Expression; D005810:Multigene Family; D009154:Mutation; D030821:Plants, Genetically Modified / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000145:classification / Q000235:genetics* / Q000502:physiology*; D020798:Two-Hybrid System Techniques
D029681:Arabidopsis Proteins; D012333:RNA, Messenger; D014157:Transcription Factors