doc. Mgr. Petra Procházková Schrumpfová, Ph.D.
Research topics
Transcription of individual genes in plant cells must be controlled very precisely. Its precise regulation is essential for optimal plant growth, the development of sexual reproductive organs, or the adaptation of plants to changing environmental conditions. The regulation of transcription appears to require the presence of various specific regulatory motifs and elements in the promoters of genes that modulate transcription through their associated proteins.
Research group member:
Our research focuses on characterizing gene regulatory sequences and their associated proteins. We pay particular attention to the telomeric repeat, found not only at the physical ends of chromosomes but also as a short repeat called a telo-box in the promoters of many genes. We are also studying proteins that recognize these regulatory sequences, such as the TRB (Telomere Repeat Binding) family of proteins and their associated proteins, which are involved in the regulation of transcription through telo-boxes and also in the protection of chromosome ends.
We use a wide range of methods and techniques from molecular biology and biochemistry (e.g., cloning techniques; methods for protein expression and purification; techniques to verify protein-protein interactions (Y2H; Co-IP; BiFC) and protein-DNA interactions (ChIP-Seq; EMSA)). We aim to link the results from these analyses to changes at the level of plant physiology - for example, results obtained by specific staining at the cellular level or detailed analysis of mutant plants for selected genes (e.g. GUS-staining; aniline-blue staining; in vitro growth of pollen tubes). We are also involved in the development of software programs used for detailed localization of the distribution of gene regulatory elements in gene promoters in different plants (GOLEM software).
Main objectives
- Characterization of proteins associated with gene regulatory sequences
- Study the role of gene regulatory elements on gene expression
- Detection of the function of telomeric repeats in gene promoters
- Localizing the distribution of gene regulatory elements near transcriptional origins
a) EMSA of protein binding to radioactively labelled oligonucleotides. b) Detection of telomerase activity. c) Interactions of proteins fused with nYFP or cYFP part were detected using the Bimolecular fluorescence complementation (BiFC) assay in benthamiana leaf epidermal cells. d) Detection of promoter activity by GUS histochemical staining.e) Aniline blue staining of pistil 1 day after pollination (DAP). f) Graph exported from program GOLEM (Gene regulatOry eLEMents; https://golem.ncbr.muni.cz) which enables users to precisely locate any motif of interest with respect to TSS or ATG.
Overview of the main Telomere repeat binding proteins (TRBs) functions. a) TRBs are associated with the physical ends of chromosomes (telomeres) and interact with POT1 proteins. b) TRBs mediate interactions of RUVBL proteins with the catalytic subunit of telomerase (TERT), and participate in telomerase biogenesis. c) TRBs are associated with short telomeric sequences (telo-boxes) in the promoters of various. d) Telo-box motifs recruit PRC2 repressive complex. e) Histone H1 prevents the invasion of H3K27me3 and TRB1 over long telomeric repeats. f) TRB proteins, as subunits of the PEAT (PWO-EPCR-ARID-TRB) complex, are involved in epigenetic regulations of chromatin.