Identification and characterization of transcription factor IIIA and ribosomal protein L5 from Arabidopsis thaliana.
Thus far, no transcription factor IIIA (TFIIIA) from higher plants has been cloned and characterized. We have cloned and characterized TFIIIA and ribosomal protein L5 from Arabidopsis thaliana. Primary sequence comparison revealed a high divergence of AtTFIIIA and a relatively high conservation of AtL5 when compared with other organisms. The AtTFIIIA cDNA encodes a protein with nine Cys(2)-His(2)-type zinc fingers, a 23 amino acid spacer between fingers 1 and 2, a 66 amino acid spacer between fingers 4 and 5, and a 50 amino acid non-finger C-terminal tail. Aside from the amino acids required for proper zinc finger folding, AtTFIIIA is highly divergent from other known TFIIIAs. AtTFIIIA can bind 5S rDNA, as well as 5S rRNA, and efficiently stimulates the transcription of an Arabidopsis 5S rRNA gene in vitro. AtL5 identity was confirmed by demonstrating that this protein binds to 5S rRNA but not to 5S rDNA. Protoplast transient expression assays with green fluorescent protein fusion proteins revealed that AtTFIIIA is absent from the cytoplasm and concentrated at several nuclear foci including the nucleolus. AtL5 protein accumulates in the nucleus, especially in the nucleolus, and is also present in the cytoplasm.
Mathieu
Olivier
O
UMR CNRS 6547 BIOMOVE, Université Blaise Pascal, 24 Avenue des Landais, 63177 Aubière Cedex, France.
Yukawa
Yasushi
Y
Prieto
José-Luis
JL
Vaillant
Isabelle
I
Sugiura
Masahiro
M
Tourmente
Sylvette
S
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018983:DNA Footprinting / Q000379:methods; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012269:Ribosomal Proteins / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D035561:Transcription Factor TFIIA / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D029681:Arabidopsis Proteins; D018076:DNA, Complementary; D004275:DNA, Ribosomal; D008164:Luminescent Proteins; D012335:RNA, Ribosomal; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; D035561:Transcription Factor TFIIA; C029742:ribosomal protein L5; D049452:Green Fluorescent Proteins; D003850:Deoxyribonuclease I