Biology subjects

Monecke, T.

Publications and source records attributed to Monecke, T..

5 recordsLinked to original sources

bioRxiv2026

Two components of the early ASH1 mRNA transport machinery undergo PUN motif-dependent liquid-liquid-phase separation

In eukaryotes, mRNA localization is a widespread mechanism of spatial gene regulation. Amongst the best-studied examples is the directional transport of ASH1 mRNA during mitosis of the budding yeast Saccharomyces cerevisiae. ASH1 mRNA is co-transcriptionally bound by the two RNA-binding proteins She2p and Loc1p and subsequently exported. In the cytoplasm, the adapter She3p binds to She2p forming an active transport complex with the myosin motor Myo4p to mediate actin-dependent transport into the daughter cell. Loc1p stays in the nucleus where it has a second function in ribosome biogenesis. In this study, we map the interaction surface of Loc1p on She2p and observe that it does not interfere with the RNA-binding interface of She2p. Analogous to a previous report on Loc1p and RNA, we could show that also Loc1p and She2p undergo liquid-liquid phase separation (LLPS). This event is caused by electrostatic interactions and can be regulated by the phosphorylation-driven alteration of She2ps oligomeric state. Furthermore, we observed that LLPS formation only requires the PUN motifs of Loc1p and that a ternary Loc1p-RNA-She2p complex also undergoes LLPS. In summary, our findings indicate that Loc1p co-transcriptionally recruits the nuclear ASH1 mRNA-She2p complex to LLPS, while the cytoplasmic Loc1p-lacking She2p-She3p-RNA transport complex does not form such LLPS.

biochemistry↗

bioRxiv2024

A direct interaction between the RNA-binding proteins Staufen and Tm1-I/C regulates oskar mRNP composition and transport

In the Drosophila female germline, oskar messenger RNA is transported on microtubules from the nurse cells to the posterior pole of the oocyte, where it is translated. Transport of oskar transcripts from the nurse cells into the oocyte requires dynein, while localization of the mRNAs within the oocyte to the posterior pole is dependent upon kinesin-1. Staufen, a dsRNA-binding protein, has been shown to bind the oskar mRNA transport complex in the oocyte and inactivate dynein; however, it remains unclear how kinesin is activated. Here, using surface plasmon resonance, nuclear magnetic resonance spectroscopy and RNA imaging within egg chambers, we demonstrate that Staufen directly interacts with Tm1, a non-canonical kinesin adaptor. This work provides a molecular explanation for the previously unclear role of Staufen in oskar mRNA localization.

molecular biology↗

bioRxiv2024

Intrinsically disordered RNA-binding motifs cooperate to catalyze RNA folding and drive phase separation

RNA-binding proteins are essential for gene regulation and the spatial organization of cells. Here, we report that the yeast ribosome biogenesis factor Loc1p is an intrinsically disordered RNA-binding protein with eight repeating positively charged, unstructured nucleic acid binding (PUN) motifs. While a single of these previously undefined motifs stabilizes folded RNAs, multiple copies strongly cooperate to catalyze RNA folding. In the presence of RNA, these multivalent PUN motifs drive phase separation. Proteome-wide searches in pro-and eukaryotes for proteins with similar arrays of PUN motifs reveal a strong enrichment in RNA-mediated processes and DNA remodeling. Thus, PUN motifs are potentially involved in a large variety of RNA-and DNA-related processes by concentrating them in membrane-less organelles. The general function and wide distribution of PUN motifs across species suggests that in an ancient "RNA world" PUN-like motifs may have supported the correct folding of early ribozymes.

biochemistry↗

bioRxiv2023

PURA Syndrome-causing mutations impair PUR-domain integrity and affect P-body association

Mutations in the human PURA gene cause the neuro-developmental PURA syndrome. In contrast to several other mono-genetic disorders, almost all reported mutations in this nucleic acid binding protein result in the full disease penetrance. In this study, we observed that patient mutations across PURA impair its previously reported co-localization with processing bodies. These mutations either destroyed the folding integrity, RNA binding or dimerization of PURA. We also solved the crystal structures of the N- and C-terminal PUR domains of human PURA and combined them with molecular dynamics simulations and NMR measurements. The observed unusually high dynamics and structural promiscuity of PURA indicated that this protein is particularly susceptible to mutations impairing its structural integrity. It offers an explanation why even conservative mutations across PURA result in the full penetrance of symptoms in patients with PURA syndrome.

molecular biology↗

bioRxiv2022

Human TRMT2A methylates different components of the translation machinery and contributes to translation fidelity

Methyl-5-uridine (m5U) is one of the most abundant RNA modifications found in cytosolic tRNA. tRNA methyltransferase 2 homolog A (hTRMT2A) is the dedicated mammalian enzyme of m5U formation at tRNA position 54. However, its RNA binding specificity and functional role in the cell are not well understood. Here we dissected structural and sequence requirements for binding and methylation of its RNA targets. Specificity of tRNA modification by TRMT2A is achieved by a combination of modest binding preference and presence of a uridine in position 54 of tRNAs. Mutational analysis together with crosslinking experiments identified a large hTRMT2A-tRNA binding surface. Furthermore, complementing hTRMT2A interactome studies revealed that TRMT2A interacts with proteins involved in RNA biogenesis. Finally, we addressed the question of the importance of TRMT2A function by showing that its knockdown reduces translation fidelity. These findings extend the role of hTRMT2A beyond tRNA modification towards a role in translation.

biochemistry↗