Analysis of Rheb in the cellular slime mold Dictyostelium discoideum: cellular localization, spatial expression and overexpression.
Dictyostelium discoideum encodes a single Rheb protein showing sequence similarity to human homologues of Rheb. The DdRheb protein shares 52 percent identity and 100 percent similarity with the human Rheb1 protein. Fluorescence of Rheb yellow fluorescent protein fusion was detected in the D. discoideum cytoplasm. Reverse transcription-polymerase chain reaction and whole-mount in situ hybridization analyses showed that rheb is expressed at all stages of development and in prestalk cells in the multicellular structures developed. When the expression of rheb as a fusion with lacZ was driven under its own promoter, the beta-galactosidase activity was seen in the prestalk cells. D. discoideum overexpressing Rheb shows an increase in the size of the cell. Treatment of the overexpressing Rheb cells with rapamycin confirms its involvement in the TOR signalling pathway.
School of Life Science, Jawaharlal Nehru University, New Delhi 110 067, India.
Pynskhem Bok
PB
Swer
Pooja
P
Bhadoriya
Shweta
S
Saran
D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D004023:Dictyostelium / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D015151:Immunoblotting; D017403:In Situ Hybridization; D008164:Luminescent Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D009479:Neuropeptides / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D015398:Signal Transduction / Q000235:genetics; D058570:TOR Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D017931:DNA Primers; D008164:Luminescent Proteins; D009479:Neuropeptides; C054562:yellow fluorescent protein, Bacteria; D058570:TOR Serine-Threonine Kinases; D020559:Monomeric GTP-Binding Proteins