title: A cytoplasmic prolyl hydroxylation and glycosylation pathway modifies Skp1 and regulates O2-dependent development in Dictyostelium.
abstract: The soil amoeba Dictyostelium is an obligate aerobe that monitors O(2) for informational purposes in addition to utilizing it for oxidative metabolism. Whereas low O(2) suffices for proliferation, a higher level is required for slugs to culminate into fruiting bodies, and O(2) influences slug polarity, slug migration, and cell-type proportioning. Dictyostelium expresses a cytoplasmic prolyl 4-hydroxylase (P4H1) known to mediate O(2)-sensing in animals, but lacks HIFalpha, a major hydroxylation target whose accumulation directly induces animal hypoxia-dependent transcriptional changes. The O(2)-requirement for culmination is increased by P4H1-gene disruption and reduced by P4H1 overexpression. A target of Dictyostelium P4H1 is Skp1, a subunit of the SCF-class of E3-ubiquitin ligases related to the VBC-class that mediates hydroxylation-dependent degradation of animal HIFalpha. Skp1 is a target of a novel cytoplasmic O-glycosylation pathway that modifies HyPro143 with a pentasaccharide, and glycosyltransferase mutants reveal that glycosylation intermediates have antagonistic effects toward P4H1 in O(2)-signaling. Current evidence indicates that Skp1 is the only glycosylation target in cells, based on metabolic labeling, biochemical complementation, and enzyme specificity studies. Bioinformatics studies suggest that the HyPro-modification pathway existed in the ancestral eukaryotic lineage and was retained in selected modern day unicellular organisms whose life cycles experience varying degrees of hypoxia. It is proposed that, in Dictyostelium and other protists including the agent for human toxoplasmosis Toxoplasma gondii, prolyl hydroxylation and glycosylation mediate O(2)-signaling in hierarchical fashion via Skp1 to control the proteome, directly via degradation rather than indirectly via transcription as found in animals.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center, 975 Northeast Tenth Street, Oklahoma City, OK 73104, USA. Cwest2@ouhsc.edu
authors.forename: Christopher M
authors.initials: CM
authors.lastname: West
authors.forename: Zhuo A
authors.initials: ZA
authors.lastname: Wang
authors.forename: Hanke
authors.initials: H
authors.lastname: van der Wel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003593:Cytoplasm / Q000201:enzymology; D004023:Dictyostelium / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D005647:Fucosyltransferases / Q000737:chemistry / Q000378:metabolism; D005700:Galactosyltransferases / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology / Q000502:physiology*; D011393:Procollagen-Proline Dioxygenase / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D015800:Protozoan Proteins / Q000235:genetics; D044843:SKP Cullin F-Box Protein Ligases / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D011393:Procollagen-Proline Dioxygenase; D044843:SKP Cullin F-Box Protein Ligases; D005647:Fucosyltransferases; D005700:Galactosyltransferases; D017351:N-Acetylglucosaminyltransferases; C023224:alpha-1,3-mannosyl-glycoprotein 2-beta-N-acetylglucosaminyltransferase; D010100:Oxygen