Molecular cloning and analysis of the yeast flocculation gene FLO1.
The DNA sequence of the flocculation gene FLO1 of Saccharomyces cerevisiae, which is located on chromosome I (Watari et al., 1989) was determined. The sequence contains a large open reading frame (ORF) of 2586 bp and codes for a protein of 862 amino acids. However, further study (genomic Southern and polymerase chain reaction analyses) indicated that the gene we cloned was not the intact FLO1 gene but a form with an approximately 2 kb deletion in the ORF region. The intact FLO1 gene was then cloned and its nucleotide sequence determined. The sequence revealed that the ORF of the intact gene is composed of 4611 bp which code for a protein of 1537 amino acids. A remarkable feature of the putative Flo1 protein is that it contains four families of repeated sequences composed of 18, 2, 3 and 3 repeats and that it has a large number of serines and threonines. In the deleted FLO1 form, a large part of these repeated sequences was missing. The N- and C-terminal regions are hydrophobic and both contain a potential membrane-spanning region, suggesting that the Flo1 protein is an integral membrane protein and a cell wall component.
Watari
J
J
Brewing Research Laboratories, Sapporo Breweries Ltd., Shizuoka, Japan.
Takata
Y
Y
Ogawa
M
M
Sahara
H
H
Koshino
S
S
Onnela
M L
ML
Airaksinen
U
U
Jaatinen
R
R
Penttilä
M
M
Keränen
S
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D005427:Flocculation; D005800:Genes, Fungal / Q000235:genetics*; D016681:Genome, Fungal; D006031:Glycosylation; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D012045:Regulatory Sequences, Nucleic Acid; D012091:Repetitive Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA
C087578:FLO1 protein, S cerevisiae; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins