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Biology subjects

Schuh, C. M.

Publications and source records attributed to Schuh, C. M..

2 recordsLinked to original sources

Tethering-mediated recruitment of pioneer factors by NuRD enables signal dependent gene activation and protects cell identity

Cell identity relies on precise control of transcription factor (TF) activity, localization and interpretation of signaling. Mitogen-activated protein kinase (MAPK) signaling regulates transcription via the ERK kinase, phosphorylating components of the Mediator complex to regulate its association with TF-bound enhancers. Here we describe two distinct forms of TF association related to signaling response, stable DNA recognition and tethering to active sites of transcription. We find that the tethering of TFs at these active sites is both essential for signaling response and cell identity. To identify tethering substrates, we focused on signaling dependent associations with the transcriptional coregulator, the Mediator, and find the NuRD complex is engaged with the Mediator at active promoters and enhancers. Here, NuRD functions as a molecular scaffold recruiting pioneer TFs like SOX2 independently of DNA sequence, enhancing ERK dependent transcription while sequestering these factors away from context inappropriate binding sites elsewhere in the genome. Balancing the recognition of a restricted spectrum of canonical DNA sites by pioneer factors while retaining them at high concentrations in the vicinity of transcription supports robust transcriptional response, ensures plasticity in differentiation and explains why these factors bind a fraction of their naturally occurring sites in any given cell type.

molecular biology↗

Modulation of the Biophysical and Biochemical Properties of Collagen by Glycation for Tissue Engineering Applications

The structural and functional properties of collagen are modulated by the presence of intramolecular and intermolecular crosslinks. Advanced Glycation End-products (AGEs) can produce intermolecular crosslinks by bonding the free amino groups of neighboring proteins. In this research, the following hypothesis is explored: The accumulation of AGEs in collagen decreases its proteolytic degradation rates while increasing its stiffness. Fluorescence Lifetime Imaging (FLIM) and Fourier-transform infrared spectroscopy (FTIR) detect biochemical changes in collagen scaffolds during the glycation process. The accumulation of AGEs increases exponentially in the collagen scaffolds as a function of Methylglyoxal (MGO) concentration by performing autofluorescence measurement and competitive ELISA. Glycated scaffolds absorb water at a much higher rate confirming the direct affinity between AGEs and interstitial water within collagen fibrils. In addition, the topology of collagen fibrils as observed by Atomic Force Microscopy (AFM) is a lot more defined following glycation. The elastic modulus of collagen fibrils decreases as a function of glycation, whereas the elastic modulus of collagen scaffolds increases. Finally, the enzymatic degradation of collagen by bacterial collagenase shows a sigmoidal pattern with a much slower degradation rate in the glycated scaffolds. This study identifies unique variations in the properties of collagen following accumulation of AGEs.

bioengineering↗