Visual screening for localized RNAs in yeast revealed novel RNAs at the bud-tip.
Several RNAs, including rRNAs, snRNAs, snoRNAs, and some mRNAs, are known to be localized at specific sites in a cell. Although methods have been established to visualize RNAs in a living cell, no large-scale visual screening of localized RNAs has been performed. In this study, we constructed a genomic library in which random genomic fragments were inserted downstream of U1A-tag sequences under a GAL1 promoter. In a living yeast cell, transcribed U1A-tagged RNAs were visualized by U1A-GFP that binds the RNA sequence of the U1A-tag. In this screening, many RNAs showed nuclear signals. Since the nuclear signals of some RNAs were not seen when the U1A-tag was connected to the 3' ends of the RNAs, it is suggested that their nuclear signals correspond to nascent transcripts on GAL1 promoter plasmids. Using this screening method, we successfully identified two novel localized mRNAs, CSR2 and DAL81, which showed bud-tip localization.
Andoh
Tomoko
T
Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, Kurokami 2-39-1, Kumamoto 860-8555, Japan. andoh@sci.kumamoto-u.ac.jp
Oshiro
Yukiko
Y
Hayashi
Sachiko
S
Takeo
Hideki
H
Tani
Tokio
T
D002467:Cell Nucleus / Q000737:chemistry; D020732:Cytogenetic Analysis / Q000379:methods*; D003593:Cytoplasm / Q000737:chemistry; D015698:Genomic Library; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D012331:RNA, Fungal / Q000032:analysis*; D016601:RNA-Binding Proteins / Q000032:analysis / Q000235:genetics; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000032:analysis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics* / Q000254:growth & development
D012331:RNA, Fungal; D016601:RNA-Binding Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear; C075339:U1A protein; D049452:Green Fluorescent Proteins