bims-micpro Biomed News
on Discovery and characterization of microproteins
Issue of 2024–11–17
four papers selected by
Thomas Farid Martínez, University of California, Irvine



  1. Comput Struct Biotechnol J. 2024 Dec;23 3771-3779
      Upstream open reading frames (uORFs) are critical cis-acting regulators of downstream gene expression. Specifically, uORFs regulate translation by disrupting translation initiation or mediating mRNA decay. We herein summarize the effects of several uORFs that regulate gene expression in microbes to illustrate the detailed mechanisms mediating uORF functions. Microbes are ideal for uORF studies because of their prompt responses to stimuli. Recent studies revealed uORFs are ubiquitous in higher eukaryotes. Moreover, they influence various physiological processes in mammalian cells by regulating gene expression, mostly at the translational level. Research conducted using rapidly evolving methods for ribosome profiling combined with protein analyses and computational annotations showed that uORFs in mammalian cells control gene expression similar to microbial uORFs, but they also have unique tumorigenesis-related roles because of their protein-encoding capacities. We briefly introduce cutting-edge research findings regarding uORFs in mammalian cells.
    Keywords:  Gene expression and regulation; Microbes; Stress response; Transcription; Translation; UORFs
    DOI:  https://doi.org/10.1016/j.csbj.2024.10.042
  2. Life Sci Alliance. 2025 Feb;pii: e202402910. [Epub ahead of print]8(2):
      Proteogenomics has enabled the detection of novel proteins encoded in noncanonical or alternative open reading frames (altORFs) in genes already coding a reference protein. Reanalysis of proteomic and ribo-seq data revealed that the p53-induced death domain-containing protein (or PIDD1) gene encodes a second 171 amino acid protein, altPIDD1, in addition to the known 910-amino acid-long PIDD1 protein. The two ORFs overlap almost completely, and the translation initiation site of altPIDD1 is located upstream of PIDD1. AltPIDD1 has more translational and protein level evidence than PIDD1 across various cell lines and tissues. In HEK293 cells, the altPIDD1 to PIDD1 ratio is 40 to 1, as measured with isotope-labeled (heavy) peptides and targeted proteomics. AltPIDD1 localizes to cytoskeletal structures labeled with phalloidin and interacts with cytoskeletal proteins. Unlike most noncanonical proteins, altPIDD1 is not evolutionarily young but emerged in placental mammals. Overall, we identify PIDD1 as a dual-coding gene, with altPIDD1, not the annotated protein, being the primary product of translation.
    DOI:  https://doi.org/10.26508/lsa.202402910
  3. BMC Genomics. 2024 Nov 09. 25(1): 1061
       BACKGROUND: The 5' untranslated region of mRNA strongly impacts the rate of translation initiation. A recent convolutional neural network (CNN) model accurately quantifies the relationship between massively parallel synthetic 5' untranslated regions (5'UTRs) and translation levels. However, the underlying biological features, which drive model predictions, remain elusive. Uncovering sequence determinants predictive of translation output may allow us to develop a more detailed understanding of translation regulation at the 5'UTR.
    RESULTS: Applying model interpretation, we extract representations of regulatory logic from CNNs trained on synthetic and human 5'UTR reporter data. We reveal a complex interplay of regulatory sequence elements, such as initiation context and upstream open reading frames (uORFs) to influence model predictions. We show that models trained on synthetic data alone do not sufficiently explain translation regulation via the 5'UTR due to differences in the frequency of regulatory motifs compared to natural 5'UTRs.
    CONCLUSIONS: Our study demonstrates the significance of model interpretation in understanding model behavior, properties of experimental data and ultimately mRNA translation. By combining synthetic and human 5'UTR reporter data, we develop a model (OptMRL) which better captures the characteristics of human translation regulation. This approach provides a general strategy for building more successful sequence-based models of gene regulation, as it combines global sampling of random sequences with the subspace of naturally occurring sequences. Ultimately, this will enhance our understanding of 5'UTR sequences in disease and our ability to engineer translation output.
    Keywords:  5’ untranslated region; Deep neural networks; Explainable artificial intelligence; Massively parallel reporter assay; Translation regulation
    DOI:  https://doi.org/10.1186/s12864-024-10925-8
  4. Methods Mol Biol. 2025 ;2875 99-110
      Recent advances in fluorescence microscopy have now made it possible to measure the translation dynamics of individual RNA in living cells and in multiple colors. Here we describe a protocol that exploits these recent advances to simultaneously image the translation of two open reading frames encoded on a single reporter RNA yet frameshifted with respect to each other. This enables precise measurements of frameshifting dynamics and efficiency from specific frameshift stimulatory sequences, all with single-RNA precision.
    Keywords:  Frameshifting; Gene regulation; Single-molecule imaging; Translation; Viruses
    DOI:  https://doi.org/10.1007/978-1-0716-4248-1_9