Autor según el artículo: Bookey, Niamh; Drago, Paola; Leung, Kit-Yi; Hughes, Linda; MacCooey, Aoife; Ozaki, Mari; Henry, Michael; De Castro, Sandra C P; Doykov, Ivan; Heywood, Wendy E; Mills, Kevin; Murphy, Michelle M; Cavalle-Busquets, Pere; Campbell, Susan; Burtenshaw, Denise; Meleady, Paula; Cahill, Paul A; Greene, Nicholas D E; Parle-McDermott, Anne
Departamento: Ciències Mèdiques Bàsiques
Autor/es de la URV: Murphy, Michelle
Palabras clave: Synthase Rat Pseudogene Nitric-oxide Mouse Double minute chromosomes Dn Dihydrofolate-reductase genes Cells Amplification
Resumen: A functional role has been ascribed to the human dihydrofolate reductase 2 ( DHFR2 ) gene based on the enzymatic activity of recombinant versions of the predicted translated protein. However, the in vivo function is still unclear. The high amino acid sequence identity (92%) between DHFR2 and its parental homolog, DHFR, makes analysis of the endogenous protein challenging. This paper describes a targeted mass spectrometry proteomics approach in several human cell lines and tissue types to identify DHFR2-specific peptides as evidence of its translation. We show definitive evidence that the DHFR2 activity in the mitochondria is in fact mediated by DHFR, and not DHFR2. Analysis of Ribo-seq data and an experimental assessment of ribosome association using a sucrose cushion showed that the two main Ensembl annotated mRNA isoforms of DHFR2, 201 and 202, are differentially associated with the ribosome. This indicates a functional role at both the RNA and protein level. However, we were unable to detect DHFR2 protein at a detectable level in most cell types examined despite various RNA isoforms of DHFR2 being relatively abundant. We did detect a DHFR2-specific peptide in embryonic heart, indicating that the protein may have a specific role during embryogenesis. We propose that the main functionality of the DHFR2 gene in adult cells is likely to arise at the RNA level.
Áreas temáticas: Química Nutrição Molecular biology Medicine (miscellaneous) Medicina iii Medicina ii Medicina i Interdisciplinar Farmacia Ciências biológicas iii Ciências biológicas ii Ciências biológicas i Ciência da computação Biotecnología Biochemistry Biochemical research methods Analytical chemistry
Acceso a la licencia de uso: https://creativecommons.org/licenses/by/3.0/es/
Direcció de correo del autor: michelle.murphy@urv.cat
Identificador del autor: 0000-0002-6304-6204
Fecha de alta del registro: 2025-02-18
Versión del articulo depositado: info:eu-repo/semantics/publishedVersion
Referencia al articulo segun fuente origial: Molecular & Cellular Proteomics. 23 (3): 100718-
Referencia de l'ítem segons les normes APA: Bookey, Niamh; Drago, Paola; Leung, Kit-Yi; Hughes, Linda; MacCooey, Aoife; Ozaki, Mari; Henry, Michael; De Castro, Sandra C P; Doykov, Ivan; Heywood, (2024). The Differential Translation Capabilities of the Human DHFR2 Gene Indicates a Developmental and Tissue-Specific Endogenous Protein of Low Abundance. Molecular & Cellular Proteomics, 23(3), 100718-. DOI: 10.1016/j.mcpro.2024.100718
URL Documento de licencia: https://repositori.urv.cat/ca/proteccio-de-dades/
Entidad: Universitat Rovira i Virgili
Año de publicación de la revista: 2024
Tipo de publicación: Journal Publications