Article ; Online: The Differential Translation Capabilities of the Human DHFR2 Gene Indicates a Developmental and Tissue-Specific Endogenous Protein of Low Abundance.
Molecular & cellular proteomics : MCP
2024 Volume 23, Issue 3, Page(s) 100718
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 ... ...
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 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. |
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MeSH term(s) | Humans ; Cell Line ; Peptides/metabolism ; Protein Biosynthesis ; Ribosomes/metabolism ; RNA/metabolism ; RNA, Messenger/metabolism ; Tetrahydrofolate Dehydrogenase/genetics ; Tetrahydrofolate Dehydrogenase/metabolism |
Chemical Substances | Peptides ; RNA (63231-63-0) ; RNA, Messenger ; Tetrahydrofolate Dehydrogenase (EC 1.5.1.3) ; dihydrofolate reductase type II (EC 1.5.1.-) |
Language | English |
Publishing date | 2024-01-14 |
Publishing country | United States |
Document type | Journal Article |
ZDB-ID | 2075924-1 |
ISSN | 1535-9484 ; 1535-9476 |
ISSN (online) | 1535-9484 |
ISSN | 1535-9476 |
DOI | 10.1016/j.mcpro.2024.100718 |
Database | MEDical Literature Analysis and Retrieval System OnLINE |
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