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Brouhard, G.

Publications and source records attributed to Brouhard, G..

2 recordsLinked to original sources

Kappa (κ): Analysis of Curvature in Biological Image Data using B-splines

Curvature is a central morphological feature of tissues, cells, and sub-cellular structures. A challenge for computational biology is to measure the curvature of these structures from biological image data. We present an open-source Fiji plugin for measuring curvature using B-splines. The plugin is named Kappa after the Greek symbol for curvature,{kappa} . Kappa is semi-automated: users create an initialization curve by a point-click method, and the initialization curve is fit to the underlying data using an iterative minimization algorithm. We demonstrate Kappas applicability on images of cytoskeletal filaments in vitro, the cell wall of budding yeast, and whole worms moving in an agar dish. In order to verify the accuracy and precision of Kappa, we created a bank of synthetic images of known curvature using sine waves and golden spirals, which we digitized with different signal-to-noise ratios (SNR), pixel sizes, and point-spread functions (PSF). For synthetic images with characteristics similar to real data, the measured curvatures of those images show a high correlation with the theoretical curvatures. Our fitting algorithms perform better with higher SNR, smaller pixel sizes, and especially PSFs equivalent to super-resolution microscopy data (surprise, surprise). Kappa is freely available under the MIT license for simple integration into Fiji-based workflows. The source code and documentation can be found on GitHub at https://github.com/brouhardlab/Kappa.

bioinformatics

Crystal structure of human PACRG in complex with MEIG1

In human, the Parkin Co-Regulated Gene (PACRG) shares a bidirectional promoter with Parkin, a gene involved in Parkinsons disease, mitochondrial quality control and inflammation. The PACRG protein is essential to the formation of the inner junction between doublet microtubules of the axoneme, a structure found in flagella and cilia. PACRG interacts with tubulin as well as the meiosis expressed gene 1 (MEIG1) protein, which is essential for spermiogenesis in mice. However, the 3D structure of human PACRG is unknown. Here, we report the crystal structure of the C-terminal domain of human PACRG in complex with MEIG1 at 2.1 [A] resolution. PACRG adopts an -helical structure with a loop insertion that mediates a conserved network of interactions with MEIG1. Using the cryo-electron tomography structure of the axonemal doublet microtubule from the flagellated protozoan Chlamydomonas reinhardtii, we generate a model of a mammalian microtubule doublet inner junction, which reveals how PACRG interacts with tubulin subunits in both the A- and B-tubules. Furthermore, the model shows that MEIG1 interacts with {beta}-tubulin on the outer surface of the B-tubule, facing towards the central pair of the axoneme. We also model the PACRG-like protein (PACRGL), a homolog of PACRG with potential roles in microtubule remodelling and axonemal inner junction formation. Finally, we explore the evolution of the PACRG and Parkin head-to-head gene structure and analyze the tissue distribution of their transcripts. Our work establishes a framework to assess the function of the PACRG family of proteins and its adaptor proteins in the function of motile and non-motile cilia.

cell biology