Biology subjects

Peterson, C. L.

Publications and source records attributed to Peterson, C. L..

11 recordsLinked to original sources

bioRxiv2026

Functional interplay between H2B ubiquitylation and H2A.Z deposition

The Bre1 ubiquitin conjugating enzyme catalyzes the monoubiquitylation of histone H2B-K120/K123 at promoter proximal nucleosomes, contributing to transcriptional regulation. These same nucleosomes are targeted by the yeast SWR1C chromatin remodeling enzyme that deposits the histone variant H2A.Z. Yeast strains that lack both Bre1 and Swr1 are inviable, indicating that together they contribute to an essential cell function. Interestingly, H2B-K123ub and H2A.Z levels are anticorrelated, and recent biochemical studies suggest a model in which H2B-K123ub inhibits SWR1C activity by blocking access to the nucleosome acidic patch. Here, we exploit recombinant nucleosomes to show that the H2A.Z deposition activity of SWR1C is strongly inhibited by H2B-K123ub. Surprisingly, the loss of H2B-K123ub does not lead to significant changes in the genomic organization of H2A.Z in cells grown in normal media, but we find that H2B-K123ub is required for a re-localization of H2A.Z from promoters to coding regions during replication stress. Together, these data indicate a complex functional interplay between H2B ubiquitylation and H2A.Z deposition.

biochemistry↗

bioRxiv2025

CFDP1 is required for histone variant H2A.Z deposition by the human SRCAP chromatin remodeling complex

Craniofacial Developmental Protein 1 (CFDP1) is a member of the evolutionarily conserved family of Bucentaur (BCNT) proteins and was originally classified as a protein required for cell survival and differentiation during tooth development. Yeast Swc5, a BCNT family member, is an essential subunit of the yeast SWR1C chromatin remodeling complex that catalyzes the deposition of histone variant H2A.Z. Direct connections between CFDP1, H2A.Z deposition, and the mammalian SWR1 homolog, Snf2-Related CREBBP Activator Protein (SRCAP), have not been identified. Here, we perform detailed biochemical reconstitution and characterization of the human SRCAP complex (SRCAP-C). We find that CFDP1 weakly interacts with SRCAP-C in a salt concentration-dependent manner. SRCAP-C purified under a high-salt condition does not co-purify with CFDP1 and is inactive in H2A.Z dimer exchange reaction, but the addition of exogeneous CFDP1 restores the H2A.Z deposition activity of SRCAP-C, demonstrating that CFDP1 is required for H2A.Z dimer exchange by SRCAP-C. We show that CFDP1 stimulates the basal ATPase activity of reconstituted SRCAP-C, suggesting a requirement for CFDP1 in regulating intrinsic catalytic ATPase activity. Consistent with this idea, CFDP1 deficiency in human induced pluripotent stem cells (hiPSCs) leads to a genome-wide reduction of H2A.Z, H3K27me3, and H3K4me3 deposition, accompanied by the upregulation of developmental genes normally marked by these modifications. Taken together, our results provide mechanistic insights into how CFDP1 regulates histone variant H2A.Z deposition by SRCAP-C. Given mutations in the SRCAP gene cause Floating-Harbor syndrome (FHS), a rare, dominant developmental disorder, our study provides an additional link between craniofacial defects and SRCAP-mediated H2A.Z deposition.

molecular biology↗

bioRxiv2022

Prediction of Electric Fields Induced by Transcranial Magnetic Stimulation in the Brain using a Deep Encoder-Decoder Convolutional Neural Network

Transcranial magnetic stimulation (TMS) is a non-invasive, effective, and safe neuromodulation technique to diagnose and treat neurological and psychiatric disorders. However, the complexity and heterogeneity of the brain composition and structure pose a challenge in accurately determining whether critical brain regions have received the right level of induced electric field. Numerical computation methods, like finite element analysis (FEA), can be used to estimate electric field distribution. However, these methods need exceedingly high computational resources and are time-consuming. In this work, we developed a deep convolutional neural network (DCNN) encoder-decoder model to predict induced electric fields, in real-time, from T1-weighted and T2-weighted magnetic resonance imaging (MRI) based anatomical slices. We recruited 11 healthy subjects and applied TMS to the primary motor cortex to measure resting motor thresholds. Head models were developed from MRIs of the subjects using the SimNIBS pipeline. Head model overall size was scaled to 20 new size scales for each subject to form a total of 231 head models. Scaling was done to increase the number of input data representing different head model sizes. Sim4Life, a FEA software, was used to compute the induced electric fields, which served as the DCNN training data. For the trained network, the peak signal to noise ratios of the training and testing data were 32.83dB and 28.01dB, respectively. The key contribution of our model is the ability to predict the induced electric fields in real-time and thereby accurately and efficiently predict the TMS strength needed in targeted brain regions.

bioengineering↗

bioRxiv2022

H2A.Z deposition by SWR1C involves multiple ATP-dependent steps

The histone variant H2A.Z is a conserved feature of nucleosomes flanking protein-coding genes. Deposition of H2A.Z requires ATP-dependent replacement of nucleosomal H2A by a chromatin remodeler related to the multi-subunit enzyme, yeast SWR1C. How these enzymes use ATP to promote this nucleosome editing reaction remains unclear. Here we use single-molecule and ensemble methodologies to identify three ATP-dependent phases in the H2A.Z deposition reaction. Real-time analysis of single nucleosome remodeling events reveals an initial, priming step that occurs after ATP addition that likely involves transient DNA unwrapping from the nucleosome. Priming is followed by rapid loss of histone H2A, which is subsequently released from the H2A.Z nucleosomal product. Surprisingly, the rates of both priming and the release of the H2A/H2B dimer are sensitive to ATP concentration. This complex reaction pathway provides multiple opportunities to regulate the timely and accurate deposition of H2A.Z at key genomic locations.

biochemistry↗

bioRxiv2021

SWR1C catalyzes H2A.Z deposition by coupling ATPase activity to the nucleosome acidic patch

The SWR1C chromatin remodeling enzyme catalyzes the ATP-dependent exchange of nucleosomal histone H2A for the histone variant H2A.Z, a key variant involved in a multitude of nuclear functions. How the 14-subunit SWR1C engages the nucleosomal substrate remains largely unknown. Studies on the ISWI, CHD1, and SWI/SNF families of chromatin remodeling enzymes have demonstrated key roles for the nucleosomal acidic patch for remodeling activity, however a role for this nucleosomal epitope in nucleosome editing by SWR1C has not been tested. Here, we employ a variety of biochemical assays to demonstrate an essential role for the acidic patch in the H2A.Z exchange reaction. Utilizing asymmetrically assembled nucleosomes, we demonstrate that the acidic patches on each face of the nucleosome are required for SWR1C-mediated dimer exchange, suggesting SWR1C engages the nucleosome in a "pincer-like" conformation, engaging both patches simultaneously. Loss of a single acidic patch results in loss of high affinity nucleosome binding and nucleosomal stimulation of ATPase activity. We identify a conserved arginine-rich motif within the Swc5 subunit that binds the acidic patch and is key for dimer exchange activity. In addition, our cryoEM structure of a Swc5-nucleosome complex suggests that promoter proximal, histone H2B ubiquitinylation may regulate H2A.Z deposition. Together these findings provide new insights into how SWR1C engages its nucleosomal substrate to promote efficient H2A.Z deposition.

biochemistry↗

bioRxiv2021

SWI/SNF senses carbon starvation with a pH-sensitive low complexity sequence

It is increasingly appreciated that intracellular pH changes are important biological signals. This motivates the elucidation of molecular mechanisms of pH-sensing. We determined that a nucleocytoplasmic pH oscillation was required for the transcriptional response to carbon starvation in S. cerevisiae. The SWI/SNF chromatin remodeling complex is a key mediator of this transcriptional response. We found that a glutamine-rich low complexity sequence (QLC) in the SNF5 subunit of this complex, and histidines within this sequence, were required for efficient transcriptional reprogramming during carbon starvation. Furthermore, the SNF5 QLC mediated pH-dependent recruitment of SWI/SNF to a model promoter in vitro. Simulations showed that protonation of histidines within the SNF5 QLC lead to conformational expansion, providing a potential biophysical mechanism for regulation of these interactions. Together, our results indicate that that pH changes are a second messenger for transcriptional reprogramming during carbon starvation, and that the SNF5 QLC acts as a pH-sensor.

molecular biology↗

bioRxiv2020

Effect of Elbow Flexion Angle on Cortical Voluntary Activation of the Non-Impaired Biceps: A 3 Days Repeated Sessions Study.

Measurement of cortical voluntary activation (VA) with transcranial magnetic stimulation (TMS) is limited by technical challenges. One challenge is the difficulty in preferential stimulation of cortical neurons projecting to the target muscle and minimal stimulation of cortical neurons projecting to antagonists. Thus, the motor evoked potential (MEP) response to TMS in the target muscle compared to its primary antagonist may be an important parameter in the assessment of cortical VA. Modulating isometric elbow angle alters the magnitude of MEPs at rest. The purpose of this study was to evaluate the effect of isometric elbow flexion-extension angle on: 1) the ratio of biceps MEP relative to the triceps MEP amplitude across a range of voluntary efforts, and 2) cortical VA. Ten non-impaired participants completed three sessions wherein VA was determined using TMS at 45{degrees}, 90{degrees} and 120{degrees} of isometric elbow flexion, and peripheral electrical stimulation at 90{degrees} of elbow flexion. The biceps/triceps MEP ratio was greater in the more flexed elbow angle (120{degrees} flexion) compared to 90{degrees} during contractions of 50% and 75% of maximum voluntary contraction. Cortical VA assessed in the more extended elbow angle (45{degrees} flexion) was lower relative to 90{degrees} elbow flexion; this effect was dependent on the biceps/triceps MEP ratio. Cortical VA was sensitive to small changes in the linearity of the voluntary torque and superimposed twitch relationship, regardless of the elbow angle. Peripheral and cortical VA measures at 90{degrees} of elbow flexion were repeatable across three days. In conclusion, although the biceps/triceps MEP ratio was increased at a more flexed elbow angle relative to 90{degrees}, there was not a corresponding difference in cortical VA. Thus, increasing the MEP ratio via elbow angle did not affect estimation of cortical VA.

neuroscience↗

bioRxiv2020

SWI/SNF antagonizes SIR heterochromatin to promote transcription of genes expressed during mitotic exit in Saccharomyces cerevisiae

Heterochromatin is a repressive, specialized chromatin structure that is central to eukaryotic transcriptional regulation and genome stability. In the budding yeast, Saccharomyces cerevisiae, heterochromatin formation requires Sir2p, Sir3p, and Sir4p, and these Sir proteins create specialized chromatin structures at telomeres and silent mating type loci. Previously, we reported that the SWI/SNF chromatin remodeling enzyme can evict Sir3 from chromatin fibers in vitro, though whether this activity contributes to the role of SWI/SNF as a transcriptional activator at euchromatic loci is unknown. Here, we characterize genetic interactions between the SIR genes (SIR2, SIR3, and SIR4) and genes encoding subunits of the chromatin remodelers SWI/SNF and INO80C, as well genes encoding the histone deacetylases Hst3 and Hst4. We find that loss of SIR genes partially rescues the growth defects of swi2, ino80, and hst3/hst4 mutants during replication stress conditions. Interestingly, partial suppression of swi2, ino80, and hst3 hst4 mutant phenotypes is due to the pseudo-diploid state of sir mutants, but a significant portion is due to more direct functional interactions. Consistent with this view, transcriptional profiling of strains lacking Swi2 or Sir3 identifies a set of genes whose expression in the M/G1 phase of the cell cycle requires SWI/SNF to antagonize the repressive impact of Sir3.

genetics↗

bioRxiv2020

Ruler elements in chromatin remodelers set nucleosome array spacing and phasing

Arrays of regularly spaced nucleosomes dominate chromatin and are often phased by alignment to reference sites like active promoters. How the distances between nucleosomes (spacing), and between phasing sites and nucleosomes are determined remains unclear, and specifically, how ATP dependent chromatin remodelers impact these features. Here, we used genome-wide reconstitution to probe how Saccharomyces cerevisiae ATP dependent remodelers generate phased arrays of regularly spaced nucleosomes. We find that remodelers bear a functional element named the ruler that determines spacing and phasing in a remodeler-specific way. We use structure-based mutagenesis to identify and tune the ruler element residing in the Nhp10 and Arp8 modules of the INO80 remodeler complex. Generally, we propose that a remodeler ruler regulates nucleosome sliding direction bias in response to (epi)genetic information. This finally conceptualizes how remodeler-mediated nucleosome dynamics determine stable steady-state nucleosome positioning relative to other nucleosomes, DNA bound factors, DNA ends and DNA sequence elements.

biochemistry↗

bioRxiv2018

SIR proteins create compact heterochromatin fibers

Heterochromatin is a silenced chromatin region essential for maintaining genomic stability and driving developmental processes. The complicated structure and dynamics of heterochromatin have rendered it difficult to characterize. In budding yeast, heterochromatin assembly requires the SIR proteins -- Sir3, believed to be the primary structural component of SIR heterochromatin, and the Sir2/4 complex, responsible for the targeted recruitment of SIR proteins and the deacetylation of lysine 16 of histone H4. Previously, we found that Sir3 binds but does not compact nucleosomal arrays. Here we reconstitute chromatin fibers with the complete complement of SIR proteins and use sedimentation velocity, molecular modeling, and atomic force microscopy to characterize the stoichiometry and conformation of SIR chromatin fibers. In contrast to previous studies, our results demonstrate that SIR arrays are highly compact. Strikingly, the condensed structure of SIR heterochromatin fibers requires both the integrity of H4K16 and an interaction between Sir3 and Sir4. We propose a model in which two molecules of Sir3 bridge and stabilize two adjacent nucleosomes, while a single Sir2/4 heterodimer binds the intervening linker DNA, driving fiber compaction.

biochemistry↗

bioRxiv2018

Transient kinetic analysis of SWR1C-catalyzed H2A.Z deposition unravels the impact of nucleosome dynamics and the asymmetry of stepwise histone exchange

The SWR1C chromatin remodeling enzyme catalyzes an ATP-dependent replacement of nucleosomal H2A with the H2A.Z variant, regulating key DNA-mediated processes, such as transcription and DNA repair. Here we investigate the transient kinetic mechanism of the histone exchange reaction employing ensemble FRET, fluorescence correlation spectroscopy (FCS), and the steady state kinetics of ATP hydrolysis. Our studies indicate that SWR1C modulates nucleosome dynamics on both the millisecond and microsecond timescales, poising the nucleosome for the dimer exchange reaction. The transient kinetic analysis of the remodeling reaction performed under single turnover conditions unraveled a striking asymmetry in the ATP-dependent replacement of nucleosomal dimers, promoted by localized DNA translocation. Taken together, our transient kinetic studies identify new intermediates and provide crucial insights into the SWR1C-catalyzed dimer exchange reaction, as well as shedding light on how the mechanics of H2A.Z deposition might contribute to transcriptional regulation in vivo.

biochemistry↗