ABSTRACT
We have determined the nucleotide sequence of 129,524 bases of yeast (Saccharomyces cerevisiae) chromosome XV. Sequence analysis revealed the presence of 59 non-overlapping open reading frames (ORFs) of length > 300 bp, three tRNA genes, four delta elements and one Ty-element. Among the 21 previously known yeast genes (36% of all ORFs in this fragment) were nucleoporin (NUP1), ras protein (RAS1), RNA polymerase III (RPC1) and elongation factor 2 (EF2). Further, 31 ORFs (53% of the total) were found to be homologous to known protein or DNA sequences, or sequence patterns. For seven ORFs (11% of the total) no homology was found. Among the most interesting protein identification in this DNA fragment are an inositol polyphosphatase, the second gene of this type found in yeast (homologous to the human OCRL gene involved in Lowe's syndrome), a new ADP ribosylation factor of the arf6 subfamily, the first protein containing three C2 domains, and an ORF similar to a Bacillus subtilis cell-cycle related protein. For each ORF detailed sequence analysis was carried out, with a full consideration of its biological function and pointing out key regions of interest for further functional analysis.
Subject(s)
Chromosomes, Fungal/genetics , Nuclear Pore Complex Proteins , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/genetics , Sequence Analysis, DNA , ADP-Ribosylation Factors , Amino Acid Sequence , Bacillus subtilis/genetics , Cloning, Molecular , DNA-Binding Proteins/genetics , Fungal Proteins/genetics , GTP-Binding Proteins/chemistry , GTP-Binding Proteins/genetics , HMGB Proteins , Humans , Membrane Proteins/genetics , Molecular Sequence Data , Molecular Structure , Nuclear Proteins/genetics , Open Reading Frames , Phosphoric Monoester Hydrolases/genetics , Proteins/genetics , RNA Polymerase III/genetics , RNA, Transfer/genetics , SOXB1 Transcription Factors , Sequence Alignment , Sequence Homology, Amino Acid , Transcription Factors , ras Proteins/geneticsABSTRACT
We have determined the nucleotide sequence of a cosmid (pIX338) containing the centromere region of yeast (Saccharomyces cerevisiae) chromosome IX. The complete nucleotide sequence of 33.8 kb was obtained by using an efficient directed sequencing strategy in combination with automated DNA sequencing on the A.L.F. DNA sequencer. Sequence analysis revealed the presence of 17 open reading frames (ORFs), four of them previously known yeast genes (sly12, pan1, sts1 and prl1), a tRNA gene and the centromere motif. Exhaustive database searches detected sequence homologues of known function for as many as 14 of the 17 ORFs. These include a mammalian tyrosine kinase substrate; the Escherichia coli cell cycle protein MinD; the human inositol polyphosphate-5-phosphatase (gene OCRL) involved in Lowe's syndrome, a developmental disorder; and helicases, for which the new yeast member defines a distinct DEAD/H-box subfamily. A surprisingly large fraction of the ORFs (at least six out of 17) in the centromeric region are apparently involved in RNA or DNA binding.
