An unusual fibrillarin gene and protein: structure and functional implications.
The diploid germinal nucleus of the ciliated protozoan Tetrahymena thermophila is unusual among eukaryotes in that it encodes a single copy of the gene for rRNA allowing identification of cis-acting mutations in rDNA affecting rRNA structure, function, and processing. The generally conserved nucleolar protein fibrillarin has been characterized from a number of systems and is involved in pre-rRNA processing. We have demonstrated that Tetrahymena has fibrillarin and have analyzed the cDNA and the genomic DNA encoding this protein. The derived amino acid sequence of the N-terminal region of Tetrahymena fibrillarin shows little similarity with the generally highly conserved glycine/arginine-rich N-terminal domain of other eukaryotic fibrillarins. The remainder of the amino acid sequence of the molecule is more conserved. Polyclonal antibodies generated against the full-length Tetrahymena fibrillarin expressed in bacteria recognize a protein of M(r) approximately 32,000 in whole-cell or nucleolar preparations. Immunocytochemistry localizes fibrillarin to nucleoli in the somatic macronuclei of vegetative cells. Transformation experiments demonstrate that fibrillarin is an essential protein in Tetrahymena. The Tetrahymena fibrillarin is expressed but does not complement a NOP1 null mutation when transformed into the yeast Saccharomyces cerevisiae, indicating less functional conservation among fibrillarins than previously suggested.
David
E
E
Department of Biology, York University, Toronto, Ontario, Canada.
McNeil
J B
JB
Basile
V
V
Pearlman
R E
RE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D004275:DNA, Ribosomal / Q000378:metabolism*; D017125:Genes, Protozoan; D008969:Molecular Sequence Data; D010802:Phylogeny; D015800:Protozoan Proteins / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012995:Solubility; D016808:Tetrahymena thermophila / Q000235:genetics*
D002868:Chromosomal Proteins, Non-Histone; D004275:DNA, Ribosomal; D015800:Protozoan Proteins; C047458:fibrillarin