Isolation and characterization of the fission yeast gene Sprpa12+ reveals that the conserved C-terminal zinc-finger region is dispensable for the function of its product.
RNA polymerase I of Saccharomyces cerevisiae contains a small subunit, A12.2, encoded by RPA12, that was previously shown to be involved in the assembly and/or stabilization of the largest subunit, A190, of RNA polymerase I. To examine whether an equivalent subunit is present in another eukaryotic RNA polymerase I, we have cloned a Schizosaccahromyces pombe cDNA that is able to complement the rpa12 mutation in S. cerevisiae. The gene, named Sprpa12+, encodes a polypeptide of 119 amino acids that shows 55% identity to S. cerevisiae A12. 2 over its entire length, including two zinc-finger motifs. Disruption of the chromosomal Sprpa12+ gene shows that it is required for growth at higher temperatures but not at lower temperatures. Expression of Sprpa190+/nuc1+, which encodes the largest subunit of the S. pombe RNA polymerase I, from a multicopy plasmid can partially suppress the growth defect of the Sprpa12 disruptant at higher temperatures. These findings suggest that A12.2 subunit is functionally and structurally conserved between S. cerevisiae and S. pombe. Finally, the analysis of mutants suggests that SpRPA12 requires the zinc-finger domain in the N-terminal region but not the one in the C-terminal region for its function.
Imazawa
Y
Y
Department of Biochemistry, Saitama Medical School, Moroyama, Japan.
Imai
K
K
Yao
Y
Y
Yamamoto
K
K
Hisatake
K
K
Muramatsu
M
M
Nogi
Y
Y
D000595:Amino Acid Sequence; D015825:Chromosomes, Fungal; D017124:Conserved Sequence; D005816:Genetic Complementation Test; D005838:Genotype; D008969:Molecular Sequence Data; D010957:Plasmids; D021122:Protein Subunits; D012318:RNA Polymerase I / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D016335:Zinc Fingers
D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; C423311:RPA12 protein, S cerevisiae; D012318:RNA Polymerase I