Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Two chitin synthases in Saccharomyces cerevisiae.

P Orlean

    The Journal of Biological Chemistry
    |April 25, 1987
    PubMed
    Summary

    Yeast has two chitin synthases: chitin synthase II, active in vivo, and chitin synthase I, a trypsin-activable zymogen. Chitin synthase II is crucial for cell wall synthesis, while chitin synthase I

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    Ynl038wp (Gpi15p) is the Saccharomyces cerevisiae homologue of human Pig-Hp and participates in the first step in glycosylphosphatidylinositol assembly.

    Yeast (Chichester, England)·2001
    Same author

    The essential Smp3 protein is required for addition of the side-branching fourth mannose during assembly of yeast glycosylphosphatidylinositols.

    The Journal of biological chemistry·2001
    Same author

    Dolichol phosphate mannose synthase from the filamentous fungus Trichoderma reesei belongs to the human and Schizosaccharomyces pombe class of the enzyme.

    Glycobiology·2000
    Same author

    Photoaffinity labelling with P3-(4-azidoanilido)uridine 5'-triphosphate identifies gpi3p as the UDP-GlcNAc-binding subunit of the enzyme that catalyses formation of GlcNAc-phosphatidylinositol, the first glycolipid intermediate in glycosylphosphatidylinositol synthesis.

    The Biochemical journal·2000
    Same author

    Glycosylphosphatidylinositol biosynthesis defects in Gpi11p- and Gpi13p-deficient yeast suggest a branched pathway and implicate gpi13p in phosphoethanolamine transfer to the third mannose.

    Molecular biology of the cell·2000
    Same author

    Congenital disorders of glycosylation caused by defects in mannose addition during N-linked oligosaccharide assembly.

    The Journal of clinical investigation·2000

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Yeast Genetics

    Background:

    • The yeast CHS1 gene encodes trypsin-activable chitin synthase I, but its disruption resulted in strains with normal chitin levels, suggesting other chitin synthases.
    • Previous studies indicated apparent absence of chitin synthase activity in vitro upon CHS1 gene disruption, despite normal in vivo chitin content.

    Purpose of the Study:

    • To investigate the presence and characteristics of other chitin synthetic activities in yeast.
    • To differentiate and characterize chitin synthase I and a second chitin synthetic activity, termed chitin synthase II.

    Main Methods:

    • Enzyme assays on disrupted (chs1 :: URA3) and wild-type yeast strains.
    • Characterization of chitin synthases based on substrate specificity (UDP-GlcNAc), product (alkali-insoluble chitin), inhibitor sensitivity (Polyoxin D), pH/temperature optima, and response to trypsin and other treatments.
    • Analysis of enzyme activity during different growth phases and in response to alpha-mating pheromone.

    Main Results:

    • Disrupted strains possess a particulate chitin synthetic activity, identified as chitin synthase II.
    • Wild-type strains contain both chitin synthase I and chitin synthase II.
    • Chitin synthase II is active without trypsin treatment, shows optimal activity in growing cultures, and is not activated by proteolysis.
    • Chitin synthase I is a trypsin-activable zymogen, detectable only after preincubation with trypsin, and its levels increase upon alpha factor treatment without affecting chitin synthesis.
    • Both enzymes utilize UDP-GlcNAc, produce alkali-insoluble chitin, and are equally sensitive to Polyoxin D, but differ in other properties.

    Conclusions:

    • Yeast possesses at least two distinct chitin synthases: chitin synthase II, likely responsible for in vivo chitin synthesis, and chitin synthase I, a zymogen possibly inactive under normal physiological conditions.
    • Chitin synthase I is not essential for alpha-mating pheromone-induced chitin synthesis, despite increased zymogen levels.

    Related Experiment Videos