Rice has two distinct classes of protein kinase genes related to SNF1 of Saccharomyces cerevisiae, which are differently regulated in early seed development.
We have isolated five cDNA clones (osk1-5) for protein kinases from rice which are related to SNF1 protein kinase of Saccharomyces cerevisiae. Based on the sequence homology, these cDNAs can be classified into two groups, group 1 (osk1) and group 2 (osk2-5). The products of these genes were demonstrated to be functional SNF1-related protein kinases by in vitro and in vivo experiments. Recombinant proteins expressed from both groups of genes were fully active as protein kinases and could phosphorylate SAMS peptide, a substrate specific for the SNF1/AMPK family, as well as themselves (autophosphorylation). Moreover, expression of osk3 cDNA in yeast snf1 mutants restored SNF1 function. Northern blot analyses showed differential expression of these two gene groups; group 1 is expressed uniformly in growing tissues (young roots, young shoots, flowers, and immature seeds), whereas group 2 is strongly expressed in immature seeds. SNF1-related protein kinases have been reported from different plant species, such as rye, barley, Arabidopsis, tobacco, and potato, while the type of gene strongly expressed in immature seeds is known only in cereals such as rye, barley, and, from our findings, in rice. Expression levels of the group 2 genes were further analyzed in seeds during seed maturation. Expression is transiently increased in the early stages of seed maturation and then decreases. The expression peak precedes those of the sbe1 and waxy genes, which are involved in starch synthesis in rice. Taken together, these findings suggest that group 2 OSK genes play important roles in the early stages of endosperm development in rice seeds.
Takano
M
M
Department of Molecular Genetics, National Institute of Agrobiological Resources, Tsukuba, Japan. mtakano@abr.affrc.go.jp
Kajiya-Kanegae
H
H
Funatsuki
H
H
Kikuchi
S
S
D050260:Carbohydrate Metabolism; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D010802:Phylogeny; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000145:classification / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D012639:Seeds / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D014018:Tissue Distribution
D012333:RNA, Messenger; D018749:RNA, Plant; C071570:SNF1-related protein kinases; D017346:Protein Serine-Threonine Kinases