Investigation of human RNA helicase DDX3X and its process of phase separation (LLPS) through mutations identified in patients with neurodevelopmental disorders

• Grant number: 22/12010-0
• Support Opportunities: Regular Research Grants
• Start date: March 01, 2023
• End date: February 28, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Juliana Ferreira de Oliveira
• Grantee: Juliana Ferreira de Oliveira
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated researchers: Ivan Rosa e Silva ; Juliana Helena Costa Smetana
• Associated scholarship(s): 24/03827-8 - Production of human protein DDX3X containing mutations identified in patients with neurodevelopmental disorders, BP.TT; 23/03919-7 - Production of human protein DDX3X containing mutations identified in patients with neurodevelopmental disorders, BP.TT

Abstract

Neurodevelopmental disorders (NDD) constitute a set of clinical conditionscharacterized by disturbances during brain development, among the most common areautism spectrum disorder (ASD) and intellectual disability (ID). To extend the comprehensionabout the molecular mechanisms related to these disorders, this research project proposesto study an important human RNA helicase from DEAD-box family, DDX3X, highly associatedwith ASD and ID, through genetic mutations identified in patients with NDDs. DDX3X isinvolved in several steps of RNA metabolism, essential for the correct neural development,and essential for liquid-liquid phase separation (LLPS), being also target for other humandiseases as cancer (leukemia, lymphoma) and viruses (HIV, dengue, SARS-CoV-2). The aimof this project is to study five missense mutations in DDX3X identified in patients with ID and ASD to analyze the impact on protein structure/function relationship. Our previous works have shown that disease-related genetic mutations are powerful tools to understand the action mechanism of important human proteins in atomic level. In this project, recombinant DDX3Xwill be produced to perform enzymatic assays, aiming to comprehend the impact of mutationsin DDX3X activity; biophysical studies, to characterize mutants and its interactions withsubstrates (RNA and ATP); structural studies using X ray crystallography, NMR and electroncryo-microscopy to investigate the mechanisms involved in protein activity; and LLPSformation in vitro and in cell by image techniques, to check if mutations interfere in this important cellular process. In summary, this project intends to increase the comprehensionabout the functioning of an important human protein associated with pathological disordersand essential for cellular phase separation process, to promote the knowledge about themolecular mechanisms related to NDDs and aiming to contribute to the future developmentof molecules that can act as chemical probes or drugs for ID and ASD. (AU)

Scientific publications

(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)

SILVA, IVAN ROSA E; SMETANA, JULIANA HELENA COSTA; DE OLIVEIRA, JULIANA FERREIRA. A comprehensive review on DDX3X liquid phase condensation in health and neurodevelopmental disorders. International Journal of Biological Macromolecules, v. 259, p. 15-pg., 2024-01-16. (22/12010-0)