Modulation of mannosylphosphodolichol synthase and dolichol kinase activity in Trichoderma, related to protein secretion

• Autorzy: Kruszewska J. , Kubicek C.P. , Palamarczyk G.
• Języki publikacji: EN

Abstrakty

It has been postulated that exoprotein secretion in Trichoderma is related to their O-glycosylation. In the present paper the involvement of phosphodolichol in this process is described and the key role of mannosylphosphodolichol (MPD) synthase in protein O-man- nosylation is discussed. The effect of water soluble phospholipid precursors such as choline and Tween 80, known also to increase secretion of cellulases when added to the medium, on MPD-synthase activity is presented. This effect is positive in the Trichoderma reesei QM 9414 (a low producing strain) but has no influence on the enzyme activity from the RUT C-30 strain selected to overproduce secretion of exoproteins and known to contain an increased cellular amount of endoplasmic reticulum. The positive effect of addition of choline and Tween to the medium on the level of dolichol kinase activity is also demonstrated. The influence of cultivation temperature on the activity of the various enzymes involved in dolichol-dependcnt protein glycosylation i.e. MPD-synthase, dolichyl kinase and MPD/ Protein mannosyl transferase was tested. For all enzymes cultivation at 35°C led to the elevated activity, which was most striking for dolichol kinase, whereas for MPD-synthase and MPD/Protein mannosyl transferase the difference was only apparent in the assay when endogenous phosphodolichol was used as a substrate. Furthermore, lipid extract from the membranes cultivated at elevated temperature, when added to the enzyme obtained from Trichoderma grown at 25°C, enhanced the dolichol kinase activity measured in the absence of exogenous dolichol. All these results suggest that the amount of endogenous dolichol as well as phosphodolichol in Trichoderma might be increased upon cultivation of the fungus at elevated temperature.

Słowa kluczowe

EN

Trichoderma; dolichol kinase; protein secretion; mannosylphosphodolichol synthase; protein kinase

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1994
• Tom: 41
• Numer: 3
• Opis fizyczny: p.331-337,fig.

Twórcy

autor

Kruszewska J.

• Polish Academy of Sciences, A.Pawinskiego 5a, 02-106 Warsaw, Poland

autor

Kubicek C.P.

autor

Palamarczyk G.

Bibliografia

• 1. Kubicck, C.P., Messner, R., Gruber, F., Mach, R. & Kubicek-Pranz, E.M. (1993) The Trichoderma cellulase regulatory puzzle: From the interior life of a secretory fungus. Enzyme Microb. Technol. 15,90-99.
• 2. Terri, T.T., Lechtovaara, P.P., Kauppinen, S., Salouvori, I. & Knowles, J. (1987) Homologous domains in Trichoderma reesei cellulytic enzymes: gene sequence and expression of cellobiohydrolase II. Gene 51,43-52.
• 3. Messner, R., Gruber, F. & Kubicck, C.P. (1988) Differential regulation of synthesis of multiple forms of specific endoglucanases by Trichodcrma reesei QM9414. J. Bacterid. 170,3689-3693.
• 4. Tomme, P., Van TUburgh, 11., Pettersson, G., Van Damme, J., Vanderckhove, J., Knowles, J., Teeri, T. & Clayessen, M. (1988) Studies of cellulolytic system of Trichoderma reesei QM 9414. Eur. /. Biochem. 170,575-581.
• 5. Salovouri, I., Makarow, M., Rauvala, H., Knowles, J. & Kaariainen, L. (1987) Low molecular weight high-mannosc lype glycans in a secreted protein of the filamentous fungus Trichoderma reesei. BiofTechnol. 5,152-156.
• 6. El-Gogary, S., Leite, A., Crivellaro, O., El Dorry, H. & Eveleigh, D.E. (1990) Tridiodenna reesei cellulase — from mutants to induction; in Trichodenna Celluloses-Biochemistry, Physiology, Genetics, and Applications (Kubicek, C.P., Eveleigh, D.E., Esterbauer, H., Steiner, W. & Kubicek-Pranz, E.M., eds.) pp. 200-210, Royal Society of Chemistry Press, Cambridge U.K.
• 7. Ghosh, A., A1 Rabiai, S., Ghosh, B.K., Trimino-Vasquez, H., Eveleigh, D.E. & Monte- necourt, 13.S. (1982) Increased endoplasmic reticulum content of a mutant of Trichoderma reesei (RUT C-30) in relation to cellulase synthesis. Enzyme Microb. Technol. 4,110-113.
• 8. Ghosh, A., Ghosh, B.K., Trimino-Vasquez, H., Eveleigh, D.E. & Montecourt, B.S. (1984) Cellulase secretion from hypercellulytic mutant of Tridiodenna reesei RUT C-30. Arch. Microbiol. 140,126-133.
• 9. Merivouri, H., Sands, J.A. & Montenecourt, B.S. (1985) Effects of tunicamycin on secretion and enzymatic activities of cellulase from Trichodentia reesei. Appl. Microbiol. Biotechnol. 23, 60-66.
• 10. Kubicek, C.P., Panda, T., Shrefer-Kunar, G., Gruber, F. & Messner, R. (1987) O-linked but not N-linked glycosylation is necessary for the secretion of endoglucanase I and II by Tricho­denna reesei. Can. J. Microbiol. 33,698-703.
• 11. I laselbech, A. & Tanner, W. (1983) O-glycosyla- tion in Sacchawmyces cerevisiae is initiated at the endoplasmic reticulum. FEBS Lett. 158,335-338.
• 12. Kruszewska, J., Messner, R., Kubicek, C.P. & Palamarczyk, G. (1989) O-glycosylation of proteins by membrane fractions of Trichodentia reesei QM 9414. /. Gen. Microbiol. 135,301-307.
• 13. Tanner, W. & Lehle, L. (1987) Protein glycosyla­tion in yeast. Biochim. Biopiiys. Acta 906,81-99.
• 14. Stahl-Bolsinger, S., Immervoll, T., Deutzmann, R. & Tanner, W. (1993) PMT1 the gene for a key enzyme of protein O-glycosylation in Saccha­wmyces cerevisiae. Prvc. Natl. Acad. Set. U.S.A. 90, 8164-8168.
• 15. Herscovics, A., Orlean, P. (1993) Glycoprotein biosynthesis in yeast. FASEB J. 7,540-550.
• 16. Orlean, P., Albright, H. & Robbins, P.W. (1988) Cloning and sequencing of the yeast gene for dolichol phosphate mannose synthase, an essential protein./. Biol. Chem. 263,17499-17507.
• 17. Banerjee, D.K., Kousvelari, E.E. & Baum, B.J. (1987) c-AMP mediated protein phosphory­lation of microsomal membranes increase mannosylphosphodolichol synthase activity. Proc. Natl. Acad. Sci. U.S.A. 84,6389-6393.
• 18. Kruszewska, J., Palamarczyk, G. & Kubicek, C.P. (1991) Mannosyl-phospho-dolichol synthase from Trichoderma reesei is activated by protein kinase dependent phosphorylation in vitro. FEMS Microbiol. Lett. 80,81-86.
• 19. Kruszewska,J., Palamarczyk,G.& Kubicck,C.P. (1990) Stimulation of exoprotein secretion by choline and Tween 80 in Trichoderma reesei QM 9414 correlates with increased activities of dolichol phosphate mannose synthase. /. Gen. Microbiol. 136,1293-1298.
• 20. Shreiber, G., Kolar, H., Foisner, R. & Kubicek, C.P. (1986) Choline stimulates synthesis of extracellular proteins in Trichodenna reesei QM 9414. Archiv. Microbiol. 144,41-47.