WFRF:(Heldin J.)
Sökning: WFRF:(Heldin J.) > (2015-2019) > Effects of mutation...
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| Fältnamn | Indikatorer | Metadata |
|---|---|---|
| 000 | 04267naa a2200385 4500 | |
| 001 | oai:DiVA.org:uu-390985 | |
| 003 | SwePub | |
| 008 | 190819s2019 | |||||||||||000 ||eng| | |
| 024 | 7 | a https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3909852 URI |
| 024 | 7 | a https://doi.org/10.1016/j.matbio.2018.10.0042 DOI |
| 040 | a (SwePub)uu | |
| 041 | a engb eng | |
| 042 | 9 SwePub | |
| 072 | 7 | a ref2 swepub-contenttype |
| 072 | 7 | a art2 swepub-publicationtype |
| 100 | 1 | a Melero-Fernandez de Mera, R. M.u Univ Eastern Finland, Inst Biomed, Kuopio, Finland4 aut |
| 245 | 1 0 | a Effects of mutations in the post-translational modification sites on the trafficking of hyaluronan synthase 2 (HAS2) |
| 264 | 1 | b Elsevier BV,c 2019 |
| 338 | a print2 rdacarrier | |
| 520 | a Vesicular trafficking of hyaluronan synthases (HAS1-3) from endoplasmic reticulum (ER) through Golgi to plasma membrane (PM), and either back to endosomes and lysosomes, or out into extracellular vesicles, is important for their activities. We studied how post-translational modifications affect the trafficking of HAS2 by mutagenesis of the sites of ubiquitination (K190R), phosphorylation (T110A) and 0-GIcNAcylation (S221A), using Dendra2- and EGFP-HAS2 transfected into COS1 cells. Confocal microscopy showed HAS2 wild type (wt) and its K19OR and S221A mutants in ER, Golgi and extracellular vesicles, while the T110A mutant remained mostly in the ER. HA synthesis was reduced by S221A, while completely blocked by K19OR and T110A. Cell-surface biotinylation indicated that T110A was absent from PM, while S221A was close to the level of wt, and K190R was increased in PM. TIRF microscopy analysis gave similar results. Rabl 0 silencing increased HA secretion by HAS2, likely by inhibiting endocytosis of the enzyme from PM, as reported before for HAS3. Green-to-red photo-conversion of Dendra2-HAS2 constructs suggested slower decay of K190R and S221A than HAS2 wt, while T110A was barely degraded at all. S221D and S221E, the phosphomimetic mutants of this site, decayed faster and blocked hyaluronan synthesis, suggesting alternative 0-GIcNAci-PO4 substitution to regulate the stability of the enzyme. Probing the role of dynamic 0-GIcNAcylation at S221 by adding glucosamine increased the half-life of only HAS2 wt. The Dendra2 " HAS2 disappearance from Golgi was slower for K190R. Of the two inactive constructs, K190R co-transfected with HAS2 wt suppressed, whereas T110A had no effect on HA synthesis. Interestingly, the HAS2stimulated shedding of extracellular vesicles was dependent on HAS residence in PM but independent of HA synthesis. The results indicate that post-translational modifications control the trafficking of HAS2, and that trafficking is an integral part of the post-translational regulation of HAS2 activity. | |
| 650 | 7 | a MEDICIN OCH HÄLSOVETENSKAPx Medicinska och farmaceutiska grundvetenskaperx Cell- och molekylärbiologi0 (SwePub)301082 hsv//swe |
| 650 | 7 | a MEDICAL AND HEALTH SCIENCESx Basic Medicinex Cell and Molecular Biology0 (SwePub)301082 hsv//eng |
| 700 | 1 | a Arasu, U. T.u Univ Eastern Finland, Inst Biomed, Kuopio, Finland4 aut |
| 700 | 1 | a Karna, R.u Univ Eastern Finland, Inst Biomed, Kuopio, Finland4 aut |
| 700 | 1 | a Oikari, S.u Univ Eastern Finland, Inst Biomed, Kuopio, Finland4 aut |
| 700 | 1 | a Rilla, K.u Univ Eastern Finland, Inst Biomed, Kuopio, Finland4 aut |
| 700 | 1 | a Vigetti, D.u Univ Insubria, Dept Med & Surg, Varese, Italy4 aut |
| 700 | 1 | a Passi, A.u Univ Insubria, Dept Med & Surg, Varese, Italy4 aut |
| 700 | 1 | a Heldin, Paraskeviu Uppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi4 aut0 (Swepub:uu)paraheld |
| 700 | 1 | a Tammi, M. I.u Univ Eastern Finland, Inst Biomed, Kuopio, Finland4 aut |
| 700 | 1 | a Deen, A. J.u Univ Eastern Finland, Inst Biomed, Kuopio, Finland;Univ Eastern Finland, AI Virtanen Inst Mol Sci, Kuopio, Finland4 aut |
| 710 | 2 | a Univ Eastern Finland, Inst Biomed, Kuopio, Finlandb Univ Insubria, Dept Med & Surg, Varese, Italy4 org |
| 773 | 0 | t Matrix Biologyd : Elsevier BVg 80, s. 85-103q 80<85-103x 0945-053Xx 1569-1802 |
| 856 | 4 8 | u https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-390985 |
| 856 | 4 8 | u https://doi.org/10.1016/j.matbio.2018.10.004 |
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