Studies conducted by researchers of BGE lab and collaborators on Drosophila melanogaster propose an important role for the Pcif1 protein in gene expression and physiology of the organism independent of its catalytic activity.

In living organisms, gene expression is a complex process that results in the production of proteins from the genome in a time- and space-regulated manner. This expression is strictly dependent on the synthesis of messenger RNAs transcripts from the DNA molecule and their subsequent translation into proteins by the ribosomes. As they are synthesized, mRNAs undergo chemical modifications that ensure their stability, their delivery to ribosomes and eventually their translation into proteins. These modifications include the addition of a cap at the 5′ end and polyadenosines tail at the 3′ end, as well as the addition of methyl groups on various nucleotides. These numerous modifications provide an additional level of information for the control of protein production and collectively form the epi-transcriptome.

In collaboration with the University of Geneva, we focused on a RNA mammalian methyltransferase, the PCIF1 protein (homologous to the Drosophila Pcif1 protein), which adds an extra methyl group to m6A (m6 adenosine) to form m6Am when the first transcribed nucleotide is an adenosine. In mice, mutation of PCIF1 encoding gene causes deregulation of the expression of a set of genes and a reduced body weight. During evolution, this protein naturally lost its catalytic activity in Drosophila where Pcif1 is, like its human counterpart, expressed in the nucleus and associated with the C-terminal domain of RNA polymerase (RNA Pol II). The work of the scientists shows that mutation of Pcif1 gene in Drosophila results in a deregulation of the expression of a set of genes, reduced body weight and a significant drop in fertility which indicates an important role for this protein in the physiology of the organism. In agreement with these observations, the Pcif1 protein is distributed all along the polytene chromosomes at the level of active transcription sites and interferes with chromatin-dependent gene expression regulation. This protein binds to the phosphorylated form of serine 5 of RNA polymerase II (Figure) and may directly modulate its activity or promote the recruitment of chromatin components. These results suggest a similar contribution of PCIF1 to the fine regulation of RNA polymerase II activity in mammals, in addition to its mRNA methylation activity, whose role in mRNA translation efficiency is actively studied but still controversial.

This study demonstrates the importance to not restrict the function of enzymes to their sole catalytic role in the living world and the interest of thoroughly examining the evolution of proteins in eukaryotes in order to discover unsuspected or hidden mechanisms of action.

The catalytic-dead Pcif1 regulates gene expression and fertility in Drosophila. Franco G, Taillebourg E, Delfino E, Homolka D, Gueguen N, Brasset E, Raman Pandey RR, Pillai RS and Fauvarque MO. RNA, 2023

The mammalian cap-specific m6Am RNA methyltransferase PCIF1 regulates transcript levels in mouse tissues. Pandey RR, Delfino E, Homolka D, Roithova A, Chen KM, Li L, Franco G, Vågbø CB, Taillebourg E, Fauvarque MO and Pillai RS. Cell Report, 2020

This work has been funded by Labex GRAL, IDEX PhD international grant and Région Rhône-Alpes