A DNA-binding protein from Candida albicans that binds to the RPG box of Saccharomyces cerevisiae and the telomeric repeat sequence of C. albicans.
Electromobility shift assays with a DNA probe containing the Saccharomyces cerevisiae ENO1 RPG box identified a specific DNA-binding protein in total protein extracts of Candida albicans. The protein, named Rbf1p (RPG-box-binding protein 1), bound to other S. cerevisiae RPG boxes, although the nucleotide recognition profile was not completely the same as that of S. cerevisiae Rap 1p (repressor-activator protein 1), an RPG-box-binding protein. The repetitive sequence of the C. albicans chromosomal telomere also competed with RPG-box binding to Rbf1p. For further analysis, we purified Rbf1p 57,600-fold from C. albicans total protein extracts, raised mAbs against the purified protein and immunologically cloned the gene, whose ORF specified a protein of 527 aa. The bacterially expressed protein showed RPG-box-binding activity with the same profile as that of the purified one. The Rbf1p, containing two glutamine-rich regions that are found in many transcription factors, showed transcriptional activation capability in S. cerevisiae and was predominantly observed in nuclei. These results suggest that Rbf1p is a transcription factor with telomere-binding activity in C. albicans.
Ishii
Nobuya
N
Department of Mycology, Nippon Roche Research Center, Kajiwara 200, Kamakura, Kanagawa-Prefecture, Japan.
Yamamoto
Mutumi
M
Department of Mycology, Nippon Roche Research Center, Kajiwara 200, Kamakura, Kanagawa-Prefecture, Japan.
Lahm
Hans-Werner
HW
Department of PRPB, F. Hoffman-La Roche AG, Postfach 4070, Basel, Switzerland.
Lizumi
Shinnji
S
Department of Mycology, Nippon Roche Research Center, Kajiwara 200, Kamakura, Kanagawa-Prefecture, Japan.
Yoshihara
Fumie
F
Department of Mycology, Nippon Roche Research Center, Kajiwara 200, Kamakura, Kanagawa-Prefecture, Japan.
Nakayama
Hironobu
H
Department of Mycology, Nippon Roche Research Center, Kajiwara 200, Kamakura, Kanagawa-Prefecture, Japan.
Arisawa
Mikio
M
Department of Mycology, Nippon Roche Research Center, Kajiwara 200, Kamakura, Kanagawa-Prefecture, Japan.
Aoki
Yuhko
Y
Department of Mycology, Nippon Roche Research Center, Kajiwara 200, Kamakura, Kanagawa-Prefecture, Japan.
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002176:Candida albicans / Q000235:genetics*; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000737:chemistry; D004271:DNA, Fungal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000235:genetics; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D015698:Genomic Library; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins; D011485:Protein Binding; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012269:Ribosomal Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017422:Sequence Analysis, DNA; D016615:Telomere / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic; D020668:rap GTP-Binding Proteins
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D009687:Nuclear Proteins; C104796:RBF1 protein, Candida albicans; D012269:Ribosomal Proteins; D014157:Transcription Factors; D019204:GTP-Binding Proteins; D020668:rap GTP-Binding Proteins