The differential translation capabilities of the human DHFR2 gene indicates a developmental and tissue specific endogenous protein of low abundance.

BOOKEY, Niamh, DRAGO, Paola, LEUNG, Kit-Yi, HUGHES, Linda, MACCOOEY, Aoife, OZAKI, Mari, HENRY, Michael, DE CASTRO, Sandra CP, DOYKOV, Ivan, HEYWOOD, Wendy E, MILLS, Kevin, MURPHY, Michelle M, CAVALLÉ-BUSQUETS, Pere, CAMPBELL, Susan, BURTENSHAW, Denise, MELEADY, Paula, CHILL, Paul A, GREENE, Nicholas and PARLE-MCDERMOTT, Anne (2024). The differential translation capabilities of the human DHFR2 gene indicates a developmental and tissue specific endogenous protein of low abundance. Molecular & cellular proteomics : MCP, 23 (3): 100718.

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Abstract

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 homologue, 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 dihydrofolate reductase 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, show differential association 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.

Item Type:	Article
Uncontrolled Keywords:	DHFR; DHFR2; Dihydrofolate Reductase; Embryogenesis; Translation; Biochemistry & Molecular Biology
Identification Number:	https://doi.org/10.1016/j.mcpro.2024.100718
SWORD Depositor:	Symplectic Elements
Depositing User:	Symplectic Elements
Date Deposited:	24 Jan 2024 13:21
Last Modified:	20 Feb 2024 13:45
URI:	https://shura.shu.ac.uk/id/eprint/33078

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