Global Identification Of Glycosylation Related Genes From Saccharomyces Ceversiae Using A Lectin Microarray

Glycosylation is an important post-translational modification thathalf of the eukaryotic proteins are glycosylated and they play importantroles in many biological processes. Recent research found thatglycosylation changes associated with tumor development and metastasis.Glycosylation depends on intracellular glycosyltransferases andtransportproteins whose alteration (knockout or overexpression) willaffect the glycosylation on cell surface. By long-term unsystematicstudies, it has found lots of glycosylation-related genes, especially insome model organisms, such as saccharomces cerevisiae. Because of thelack of biotechnology, so far no research about overall gene discovery isreported, and it is likely that there are still a significant amount ofglycosylation-related genes to be discovered.Lectin microarray provides a versatile platform for rapidhigh-throughput analysis of glycoforms of biological samples. In thisstudy we developed a high-throughput method analysing living yeast cellsurface glycans variation through profiling153strains which had arelationship with glycosylation. The result indicated the feasibility of ourmethod and it further confirmed by the results of colony PCR and MS. Base on these we profiling all strains of the knockout collections ofBY4742and the analysis of data is in process. The preliminary resultsuggests that three lectins NPL VGA and ASA had different between thestrains which was knockout the gene VAN and YLR358C respectively andwide-type. The gene VAN was expressed component of the mannanpolymerase I while YLR358C was expressed a protein of unknownfunction which may be a new gene related to glycosylation. We alsofound other four candidates through the double mutation experiment withALG5and ALG8.In this study a letin microarray was constructed and applied for forhigh-throughput profiling of yeast cell surface glycome which found fivecandidate new genes related to glycosylation. And it deserved to expectmore new discoveries with the data of microarrays analysing...