Mucor rouxii Rho1 protein; characterization and possible role in polarized growth

We have previously shown that protein kinase A of the medically important zygomycete Mucor rouxii participates in fungal morphology through cytoskeletal organization. As a first step towards finding the link between protein kinase A and cytoskeletal organization we here demonstrate the cloning of th...

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Autores principales:	Argimón, S., Galello, F., Pereyra, E., Rossi, S., Moreno, S.
Formato:	JOUR
Materias:	Caspofungin Cloning Gene expression morphology Mucor rouxii Rho1 1,3 beta glucan synthase beta 1,3 glucan caspofungin cyclic AMP dependent protein kinase fungal protein messenger RNA protein Rho1 RhoA guanine nucleotide binding protein serine unclassified drug adenosine diphosphate ribosylation amino acid sequence antifungal activity article cell differentiation controlled study correlation function culture medium enzyme active site enzyme phosphorylation enzyme substrate fungal cell fungal gene fungal morphology fungus culture fungus growth fungus mutant gene overexpression immunofluorescence test molecular cloning Mucor mucor rouxii nonhuman nucleotide sequence prediction priority journal protein analysis protein function protein localization Saccharomyces cerevisiae sequence homology Western blotting Amino Acid Sequence Base Sequence beta-Glucans Cloning, Molecular Fungal Proteins Gene Expression Regulation, Fungal Molecular Sequence Data Phylogeny Amylomyces rouxii Fungi Mammalia Zygomycetes
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00036072_v91_n3_p237_Argimon
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_00036072_v91_n3_p237_Argimon
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spelling	todo:paper_00036072_v91_n3_p237_Argimon2023-10-03T13:56:30Z Mucor rouxii Rho1 protein; characterization and possible role in polarized growth Argimón, S. Galello, F. Pereyra, E. Rossi, S. Moreno, S. Caspofungin Cloning Gene expression morphology Mucor rouxii Rho1 1,3 beta glucan synthase beta 1,3 glucan caspofungin cyclic AMP dependent protein kinase fungal protein messenger RNA protein Rho1 RhoA guanine nucleotide binding protein serine unclassified drug adenosine diphosphate ribosylation amino acid sequence antifungal activity article cell differentiation controlled study correlation function culture medium enzyme active site enzyme phosphorylation enzyme substrate fungal cell fungal gene fungal morphology fungus culture fungus growth fungus mutant gene overexpression immunofluorescence test molecular cloning Mucor mucor rouxii nonhuman nucleotide sequence prediction priority journal protein analysis protein function protein localization Saccharomyces cerevisiae sequence homology Western blotting Amino Acid Sequence Base Sequence beta-Glucans Cloning, Molecular Fungal Proteins Gene Expression Regulation, Fungal Molecular Sequence Data Mucor Phylogeny Amylomyces rouxii Fungi Mammalia Saccharomyces cerevisiae Zygomycetes We have previously shown that protein kinase A of the medically important zygomycete Mucor rouxii participates in fungal morphology through cytoskeletal organization. As a first step towards finding the link between protein kinase A and cytoskeletal organization we here demonstrate the cloning of the Rho1 gene and the characterization of its protein product. The RHO1 protein primary sequence shows 70-85% identity with fungal RHO1 or mammalian RhoA. Two protein kinase A phosphorylation sequences in adequate context are predicted, Ser73 and Ser135. The peptide IRRNSQKFV, containing Ser135 proved to be a good substrate for M. rouxii protein kinase A catalytic subunit. The over-expressed Rho1 fully complements a Saccharomyces cerevisiae null mutant. The endogenous protein was identified by western blot against a developed antibody and by ADP-ribosylation. Localization in germlings was visualized by immunofluorescence; the protein was localized in patches in the mother cell surface and excluded from the germ tube. Measurement of Rho1 expression during germination indicates that Rho1, at both the mRNA and protein levels, correlates with differentiation and not with growth. Rho1 has been shown to be the regulatory protein of the β-1,3-glucan synthase complex in fungi in which β-1,3-glucans are major components of the cell wall. Even though glucans have not been detected in zygomycetes, caspofungin, an echinochandin known to be an inhibitor of β-1,3-glucan synthase complex, is shown here to have a negative effect on growth and to produce an alteration on morphology when added to M. rouxii growth culture medium. This result has an important impact on the possible participation of β-1,3-glucans on the regulation of morphology of zygomycetes. © Springer Science+Business Media B.V. 2006. Fil:Argimón, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Galello, F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Pereyra, E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Rossi, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Moreno, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00036072_v91_n3_p237_Argimon
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Caspofungin Cloning Gene expression morphology Mucor rouxii Rho1 1,3 beta glucan synthase beta 1,3 glucan caspofungin cyclic AMP dependent protein kinase fungal protein messenger RNA protein Rho1 RhoA guanine nucleotide binding protein serine unclassified drug adenosine diphosphate ribosylation amino acid sequence antifungal activity article cell differentiation controlled study correlation function culture medium enzyme active site enzyme phosphorylation enzyme substrate fungal cell fungal gene fungal morphology fungus culture fungus growth fungus mutant gene overexpression immunofluorescence test molecular cloning Mucor mucor rouxii nonhuman nucleotide sequence prediction priority journal protein analysis protein function protein localization Saccharomyces cerevisiae sequence homology Western blotting Amino Acid Sequence Base Sequence beta-Glucans Cloning, Molecular Fungal Proteins Gene Expression Regulation, Fungal Molecular Sequence Data Mucor Phylogeny Amylomyces rouxii Fungi Mammalia Saccharomyces cerevisiae Zygomycetes
spellingShingle	Caspofungin Cloning Gene expression morphology Mucor rouxii Rho1 1,3 beta glucan synthase beta 1,3 glucan caspofungin cyclic AMP dependent protein kinase fungal protein messenger RNA protein Rho1 RhoA guanine nucleotide binding protein serine unclassified drug adenosine diphosphate ribosylation amino acid sequence antifungal activity article cell differentiation controlled study correlation function culture medium enzyme active site enzyme phosphorylation enzyme substrate fungal cell fungal gene fungal morphology fungus culture fungus growth fungus mutant gene overexpression immunofluorescence test molecular cloning Mucor mucor rouxii nonhuman nucleotide sequence prediction priority journal protein analysis protein function protein localization Saccharomyces cerevisiae sequence homology Western blotting Amino Acid Sequence Base Sequence beta-Glucans Cloning, Molecular Fungal Proteins Gene Expression Regulation, Fungal Molecular Sequence Data Mucor Phylogeny Amylomyces rouxii Fungi Mammalia Saccharomyces cerevisiae Zygomycetes Argimón, S. Galello, F. Pereyra, E. Rossi, S. Moreno, S. Mucor rouxii Rho1 protein; characterization and possible role in polarized growth
topic_facet	Caspofungin Cloning Gene expression morphology Mucor rouxii Rho1 1,3 beta glucan synthase beta 1,3 glucan caspofungin cyclic AMP dependent protein kinase fungal protein messenger RNA protein Rho1 RhoA guanine nucleotide binding protein serine unclassified drug adenosine diphosphate ribosylation amino acid sequence antifungal activity article cell differentiation controlled study correlation function culture medium enzyme active site enzyme phosphorylation enzyme substrate fungal cell fungal gene fungal morphology fungus culture fungus growth fungus mutant gene overexpression immunofluorescence test molecular cloning Mucor mucor rouxii nonhuman nucleotide sequence prediction priority journal protein analysis protein function protein localization Saccharomyces cerevisiae sequence homology Western blotting Amino Acid Sequence Base Sequence beta-Glucans Cloning, Molecular Fungal Proteins Gene Expression Regulation, Fungal Molecular Sequence Data Mucor Phylogeny Amylomyces rouxii Fungi Mammalia Saccharomyces cerevisiae Zygomycetes
description	We have previously shown that protein kinase A of the medically important zygomycete Mucor rouxii participates in fungal morphology through cytoskeletal organization. As a first step towards finding the link between protein kinase A and cytoskeletal organization we here demonstrate the cloning of the Rho1 gene and the characterization of its protein product. The RHO1 protein primary sequence shows 70-85% identity with fungal RHO1 or mammalian RhoA. Two protein kinase A phosphorylation sequences in adequate context are predicted, Ser73 and Ser135. The peptide IRRNSQKFV, containing Ser135 proved to be a good substrate for M. rouxii protein kinase A catalytic subunit. The over-expressed Rho1 fully complements a Saccharomyces cerevisiae null mutant. The endogenous protein was identified by western blot against a developed antibody and by ADP-ribosylation. Localization in germlings was visualized by immunofluorescence; the protein was localized in patches in the mother cell surface and excluded from the germ tube. Measurement of Rho1 expression during germination indicates that Rho1, at both the mRNA and protein levels, correlates with differentiation and not with growth. Rho1 has been shown to be the regulatory protein of the β-1,3-glucan synthase complex in fungi in which β-1,3-glucans are major components of the cell wall. Even though glucans have not been detected in zygomycetes, caspofungin, an echinochandin known to be an inhibitor of β-1,3-glucan synthase complex, is shown here to have a negative effect on growth and to produce an alteration on morphology when added to M. rouxii growth culture medium. This result has an important impact on the possible participation of β-1,3-glucans on the regulation of morphology of zygomycetes. © Springer Science+Business Media B.V. 2006.
format	JOUR
author	Argimón, S. Galello, F. Pereyra, E. Rossi, S. Moreno, S.
author_facet	Argimón, S. Galello, F. Pereyra, E. Rossi, S. Moreno, S.
author_sort	Argimón, S.
title	Mucor rouxii Rho1 protein; characterization and possible role in polarized growth
title_short	Mucor rouxii Rho1 protein; characterization and possible role in polarized growth
title_full	Mucor rouxii Rho1 protein; characterization and possible role in polarized growth
title_fullStr	Mucor rouxii Rho1 protein; characterization and possible role in polarized growth
title_full_unstemmed	Mucor rouxii Rho1 protein; characterization and possible role in polarized growth
title_sort	mucor rouxii rho1 protein; characterization and possible role in polarized growth
url	http://hdl.handle.net/20.500.12110/paper_00036072_v91_n3_p237_Argimon
work_keys_str_mv	AT argimons mucorrouxiirho1proteincharacterizationandpossibleroleinpolarizedgrowth AT galellof mucorrouxiirho1proteincharacterizationandpossibleroleinpolarizedgrowth AT pereyrae mucorrouxiirho1proteincharacterizationandpossibleroleinpolarizedgrowth AT rossis mucorrouxiirho1proteincharacterizationandpossibleroleinpolarizedgrowth AT morenos mucorrouxiirho1proteincharacterizationandpossibleroleinpolarizedgrowth
_version_	1807316675308027904

Mucor rouxii Rho1 protein; characterization and possible role in polarized growth

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