Biology subjects

Jang, G.-J.

Publications and source records attributed to Jang, G.-J..

3 recordsLinked to original sources

bioRxiv2024

SlimVar: rapid in vivo single-molecule tracking of chromatin regulators in plants

Epigenetic regulation occurs over many rounds of cell division in higher organisms. However, visualisation of the regulators in vivo is limited by imaging dynamic molecules deep in tissue. We report a technology--Variable-angle Slimfield microscopy (SlimVar)-- that enables tracking of single fluorescent reporters to 30 {micro}m depth through multiple Arabidopsis thaliana root tip cell layers. SlimVar uses rapid photobleaching to resolve tracked particles to molecular steps in intensity. By modifying widefield microscopy to minimise optical aberrations and robustly post-process few-photon signals, SlimVar mitigates performance losses at depth. We use SlimVar to quantify chromatin-protein assemblies in nuclei, finding that two homologous proteins key to epigenetic switching at FLOWERING LOCUS C (FLC) --cold-induced VERNALISATION INSENSITIVE3 (VIN3) and constitutively expressed VERNALISATION 5 (VRN5)--exhibit dynamic assemblies during FLC silencing. Upon cold exposure, the number of assembly molecules increases up to 100% to a median of [~]20 molecules. Larger VRN5 assemblies preferentially colocalise with an FLC lacO transgenic reporter during prolonged cold and persist after return to warmth. Our findings support a hybrid model of epigenetic memory in which nucleation of histone trimethylation is assisted by dynamic protein assemblies over extended durations. SlimVar offers molecular insights into proteins expressed at physiological levels in tissues.

plant biology↗

bioRxiv2024

Modular structure of RNA 3' processing condensates involving the Arabidopsis RNA binding protein FCA

Our understanding of the functional requirements underpinning biomolecular condensation in vivo is still relatively poor. The Arabidopsis RNA binding protein FCA is found in liquid-like nuclear condensates that function in transcription termination, promoting proximal polyadenylation at many targets in the Arabidopsis genome. To further understand the properties of these condensates in vivo we used single particle tracking experiments on FCA reporters stably expressed at endogenous levels in plant nuclei. These revealed FCA forms a core oligomer of [~]4 molecules that multimerizes into higher-order particles. This assembles into macromolecular condensates through the function of the coiled-coil protein FLL2, which is genetically required for FCA function. FLL2 predominately co-localizes with FCA in larger sized condensates. A missense mutation in the FCA RRM domain, also genetically required for FCA function, reduced both average FCA particle and condensate size but did not perturb the core oligomer. Our work points to a modular structure for FCA condensates involving multimerization of core oligomers that assemble into functional macromolecular condensates via associated RNA and FLL2 interactions.

molecular biology↗

bioRxiv2024

Functional specialization of Arabidopsis VEL polymerization domains in the switch to Polycomb silencing

Cold-induced epigenetic silencing of Arabidopsis FLOWERING LOCUS C (FLC) requires the Polycomb Repressive Complex 2 and accessory proteins VIN3 and VRN5. VIN3 and VRN5 interact via head-to-tail VEL polymerization domains, but how these functionally contribute to the switch to an epigenetically silenced state remains poorly understood. Here, we determine that VIN3 VEL polymerization involves higher order nuclear VIN3 assemblies in vivo, promotes strong chromatin association and efficient H3K27me3 nucleation. However, we also show that the polymerization domains of VIN3 and VRN5 are not equivalent: VRN5 VEL domain is not required for silencing despite its role in physically connecting VIN3 with the PRC2 complex and VRN5 VEL is unable to functionally replace VIN3 VEL in vivo. Both VIN3 and VRN5 homologs are present throughout angiosperm species, suggesting a functional requirement for maintaining different polymerization modalities. This work reveals distinct roles for multifunctional polymerization domains of Polycomb accessory proteins underpinning the onset of epigenetic silencing.

plant biology↗