Studies of the role of phosphorylation and protein binding partners in regulating the function of human class V myosins

• Grant number: 14/09720-9
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2014
• End date: March 31, 2017
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Mário Tyago Murakami
• Grantee: Mário Tyago Murakami
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated researchers: Priscila Oliveira de Giuseppe

Abstract

In this next project we will continue our studies regarding the class V myosins (MyoVs). The combination of structural and functional approaches will be maintained as in the previous project and our goal will be to answer new questions that arose during these two years of work. First, we want to know what the role of phosphorylation in regulating the CBD function of mammalian MyoVs. To this end, we intend to solve the structure of the phospho-mimmetic form of MyoVa-CBD and investigate whether phosphorylation interferes with the auto-inhibition mechanism of this molecular motor and test if it determines the selectivity of partner proteins. Our second question is about the role of alternative splicing in the recognition and selectivity of molecular partners by MyoVs. In this case, we will employ yeast two hybrid assays using as bait constructs containing the MyoVa-CBD fused to different combinations of exons. In parallel, we will perform the same test using as bait constructs of MyoVb and MyoVc similar to those built for MyoVa in order to obtain information regarding redundancy and functional divergence between these three paralogous proteins. To better understand the structural determinants for the recognition of partner proteins by MyoVs, we plan to study complexes between Rab proteins ( Ras -like small GTPases ) 3a, 8a, 10 and 39b and MyoV by X-ray crystallography , SAXS and other techniques that may be necessary. Such interactions have recently been identified in Lindsay et al (Lindsay, et al. 2013). Interestingly, Rab3a and 39b interact with the MyoVa-CBD while Rab8a and Rab10 bind to specific regions of the coiled-coil domain that undergo alternative splicing (Lindsay, et al. 2013). This set of information will have a major contribution to the understanding of structural and functional aspects of the class V myosins. (AU)

Scientific publications

(7)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

TUROWSKI, VALERIA R.; RUIZ, DIEGO M.; NASCIMENTO, ANDREY F. Z.; MILLAN, CLAUDIA; SAMMITO, MASSIMO D.; JUANHUIX, JUDITH; CREMONESI, ALINE SAMPAIO; USON, ISABEL; GIUSEPPE, PRISCILA O.; MURAKAMI, MARIO T.. Structure of the class XI myosin globular tail reveals evolutionary hallmarks for cargo recognition in plants. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, v. 77, n. 4, p. 522-533, APR 1 2021. (14/09720-9, 13/08904-6)

DOLCE, LUCIANO G.; OHBAYASHI, NORIHIKO; DA SILVA, DANIEL F. C.; FERRARI, ALLAN J. R.; PIROLLA, RENAN A. S.; SCHWARZER, ANA C. DE A. P.; ZANPHORLIN, LETICIA M.; CABRAL, LUCELIA; FIORAMONTE, MARIANA; RAMOS, CARLOS H. I.; et al. Unveiling the interaction between the molecular motor Myosin Vc and the small GTPase Rab3A. JOURNAL OF PROTEOMICS, v. 212, FEB 10 2020. (14/09720-9, 16/13195-2, 14/00584-5)

SANTOS, CLELTON A.; ZANPHORLIN, LET-CIA M.; CRUCELLO, ALINE; TONOLI, CELISA C. C.; RULLER, ROBERTO; HORTA, MARIA A. C.; MURAKAMI, MARIO T.; DE SOUZA, ANETE PEREIRA. Crystal structure and biochemical characterization of the recombinant ThBgl, a GH1 beta-glucosidase overexpressed in Trichoderma harzianum under biomass degradation conditions. BIOTECHNOLOGY FOR BIOFUELS, v. 9, MAR 22 2016. (14/09720-9, 08/52197-4, 14/18856-1, 13/13309-0)

DOLCE, LUCIANO G.; SILVA-JUNIOR, RUI M. P.; ASSIS, LEANDRO H. P.; NASCIMENTO, ANDREY F. Z.; ARAUJO, JACKELINE S.; MESCHEDE, INGRID P.; ESPREAFICO, ENILZA M.; DE GIUSEPPE, PRISCILA O.; MURAKAMI, MARIO T.. Myosin Va interacts with the exosomal protein spermine synthase. BIOSCIENCE REPORTS, v. 39, p. 11-pg., 2019-03-29. (14/00584-5, 18/04017-9, 09/14257-8, 14/03989-6, 14/09720-9, 11/20229-7, 16/10862-8, 13/08135-2)

DOLCE, LUCIANO G.; SILVA-JUNIOR, RUI M. P.; ASSIS, LEANDRO H. P.; NASCIMENTO, ANDREY F. Z.; ARAUJO, JACKELINE S.; MESCHEDE, INGRID P.; ESPREAFICO, ENILZA M.; DE GIUSEPPE, PRISCILA O.; MURAKAMI, MARIO T.. Myosin Va interacts with the exosomal protein spermine synthase. BIOSCIENCE REPORTS, v. 39, n. 3, MAR 29 2019. (18/04017-9, 14/09720-9, 16/10862-8, 14/03989-6, 13/08135-2, 09/14257-8, 14/00584-5, 11/20229-7)

ASSIS, L. H. P.; SILVA-JUNIOR, R. M. P.; DOLCE, L. G.; ALBORGHETTI, M. R.; HONORATO, R. V.; NASCIMENTO, A. F. Z.; MELO-HANCHUK, T. D.; TRINDADE, D. M.; TONOLI, C. C. C.; SANTOS, C. T.; et al. The molecular motor Myosin Va interacts with the cilia-centrosomal protein RPGRIP1L. SCIENTIFIC REPORTS, v. 7, MAR 7 2017. (14/09720-9, 14/18189-5, 09/54014-7, 11/20229-7, 09/08312-6)

TUROWSKI, VALERIA R.; RUIZ, DIEGO M.; NASCIMENTO, ANDREY F. Z.; MILLAN, CLAUDIA; SAMMITO, MASSIMO D.; JUANHUIX, JUDITH; CREMONESI, ALINE SAMPAIO; USON, ISABEL; GIUSEPPE, PRISCILA O.; MURAKAMI, MARIO T.. Structure of the class XI myosin globular tail reveals evolutionary hallmarks for cargo recognition in plants. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, v. 77, p. 12-pg., 2021-04-01. (14/09720-9, 13/08904-6)