PubMed:9613572 JSONTXT

{"project":"LocText","denotations":[{"id":"T1","span":{"begin":59,"end":68},"obj":"go:GO:0005618"},{"id":"T2","span":{"begin":81,"end":105},"obj":"taxonomy:4932"},{"id":"T3","span":{"begin":144,"end":168},"obj":"taxonomy:4932"},{"id":"T4","span":{"begin":405,"end":414},"obj":"go:GO:0005576"},{"id":"T5","span":{"begin":673,"end":682},"obj":"go:GO:0005576"},{"id":"T6","span":{"begin":691,"end":710},"obj":"uniprot:Q09187"},{"id":"T7","span":{"begin":717,"end":730},"obj":"uniprot:uniprot:"},{"id":"T8","span":{"begin":732,"end":734},"obj":"uniprot:uniprot:"},{"id":"T9","span":{"begin":806,"end":815},"obj":"go:GO:0005618"},{"id":"T10","span":{"begin":887,"end":896},"obj":"go:GO:0005618"},{"id":"T11","span":{"begin":977,"end":996},"obj":"uniprot:Q09187"},{"id":"T12","span":{"begin":1015,"end":1022},"obj":"go:GO:0009986"},{"id":"T13","span":{"begin":1090,"end":1098},"obj":"go:GO:0016020"},{"id":"T14","span":{"begin":1117,"end":1162},"obj":"uniprot:P32383"},{"id":"T15","span":{"begin":1164,"end":1170},"obj":"uniprot:P32383"},{"id":"T16","span":{"begin":1217,"end":1226},"obj":"go:GO:0005618"},{"id":"T17","span":{"begin":1252,"end":1264},"obj":"uniprot:P53753"},{"id":"T18","span":{"begin":1310,"end":1319},"obj":"go:GO:0005618"},{"id":"T19","span":{"begin":1501,"end":1510},"obj":"go:GO:0005618"}],"relations":[{"id":"R1","pred":"localizeTo","subj":"T6","obj":"T9"},{"id":"R2","pred":"localizeTo","subj":"T7","obj":"T9"},{"id":"R3","pred":"localizeTo","subj":"T7","obj":"T13"},{"id":"R4","pred":"localizeTo","subj":"T7","obj":"T16"},{"id":"R5","pred":"localizeTo","subj":"T8","obj":"T9"},{"id":"R6","pred":"localizeTo","subj":"T8","obj":"T13"},{"id":"R7","pred":"localizeTo","subj":"T8","obj":"T16"},{"id":"R8","pred":"localizeTo","subj":"T11","obj":"T10"},{"id":"R9","pred":"localizeTo","subj":"T11","obj":"T13"},{"id":"R10","pred":"localizeTo","subj":"T11","obj":"T16"},{"id":"R11","pred":"localizeTo","subj":"T11","obj":"T12"}],"namespaces":[{"prefix":"uniprot","uri":"http://identifiers.org/uniprot/"},{"prefix":"taxonomy","uri":"http://identifiers.org/taxonomy/"},{"prefix":"go","uri":"http://identifiers.org/go/"}],"text":"Screening for glycosylphosphatidylinositol (GPI)-dependent cell wall proteins in Saccharomyces cerevisiae.\nOpen reading frames in the genome of Saccharomyces cerevisiae were screened for potential glycosylphosphatidylinositol (GPI)-attached proteins. The identification of putative GPI-attached proteins was based on three criteria: the presence of a GPI-attachment signal sequence, a signal sequence for secretion and a serine- or threonine-rich sequence. In all, 53 ORFs met these three criteria and 38 were further analyzed as follows. The sequence encoding the 40 C-terminal amino acids of each was fused with the structural gene for a reporter protein consisting of a secretion signal, alpha-galactosidase and a hemagglutinin (HA) epitope, and examined for the ability to become incorporated into the cell wall. On this basis, 14 of fusion proteins were classified as GPI-dependent cell wall proteins because cells expressing these fusion proteins: (i) had high levels of alpha-galactosidase activity on their surface; (ii) released significant amounts of the fusion proteins from the membrane on treatment with phosphatidylinositol-specific phospholipase C (PI-PLC); and (iii) released fusion proteins from the cell wall following treatment with laminarinase. Of the 14 identified putative GPI-dependent cell wall proteins, 12 had novel ORFs adjacent to their GPI-attachment signal sequence. Amino acid sequence alignment of the C-terminal sequences of the 12 ORFs, together with those of known cell wall proteins, reveals some sequence similarities among them."}