Biology subjects

Walker, M. F.

Publications and source records attributed to Walker, M. F..

4 recordsLinked to original sources

bioRxiv2023

A Ciliary SMOOTHENED-GRK2-PKA Signaling Pathway Initiates Hedgehog Signal Transduction

During Hedgehog (Hh) signal transduction in development and disease, the atypical G protein-coupled receptor (GPCR) SMOOTHENED (SMO) communicates with GLI transcription factors by binding the protein kinase A catalytic subunit (PKA-C) and physically blocking its enzymatic activity. Here we show that GPCR kinase 2 (GRK2) orchestrates this process during endogenous Hh pathway activation in the vertebrate primary cilium. Upon SMO activation, GRK2 rapidly relocalizes from the ciliary base to the shaft, triggering SMO phosphorylation and PKA-C interaction. Reconstitution studies reveal that GRK2 phosphorylation enables active SMO to bind PKA-C directly. Lastly, the SMO-GRK2-PKA pathway underlies Hh signal transduction in a range of cellular and in vivo models. Thus, GRK2 phosphorylation of ciliary SMO, and the ensuing PKA-C binding and inactivation, are critical initiating events for the intracellular steps in Hh signaling. More broadly, our study suggests an expanded role for GRKs in enabling direct GPCR interactions with diverse intracellular effectors.

cell biology↗

bioRxiv2020

Species-Specific Responses of Bird Song Output in the Presence of Drones

Drones are now widely used to study wildlife, but applications for studying bioacoustics have been limited. Drones can be used to collect data on bird vocalizations, but an ongoing concern is that noise from the drones could change bird vocalization behavior. To test this behavioral impact we conducted an experiment using 30 sound localization arrays to track the song output of seven songbird species before, during, and after a 3-minute flight of a small quadcopter drone hovering at 50 m above ground level. We analyzed 8,303 song bouts, of which 2,285 song bouts of 184 individual birds were within 50 meters of the array centers. We used linear mixed effect models to assess patterns in song output showed patterns that could be attributed to the drones presence. We found no evidence of any effect of the drone for five species: American Robin Turdus migratorius, Common Yellowthroat Geothlypis trichas, Field Sparrow Spizella pusilla, Song Sparrow Melospiza melodia, and Indigo Bunting Passerina cyanea. However, we found a substantial decrease in Yellow Warbler Setophaga petechia song detections during the 3-minute drone hover, such that there was an 81% drop in detections in the 3rd minute (Wald-test, p<0.001), compared with before the drones introduction. In contrast, the number of singing Northern Cardinal Cardinalis cardinalis increased after the drone was introduced, and remained almost five-fold higher for 4-minutes after the drone departed (P<0.001). Further, we found an increase in cardinal contact/alarm calls when the drone was overhead, with the elevated calling-rate sustaining for 2 minutes after the drone had departed (P<0.001). Our study suggests that responses of songbirds to drones may be species-specific, an important consideration when proposing the use of drones in avian studies. We note that recent advances in drone technology have resulted in much quieter drones, which makes us hopeful that the impacts that we detected could be greatly reduced.

ecology↗

bioRxiv2020

Smoothened Transduces Hedgehog Signals via Activity-Dependent Sequestration of PKA Catalytic Subunits

ABSTRACTThe Hedgehog (Hh) pathway is essential for organ development, homeostasis, and regeneration. Dysfunction of this cascade drives several cancers. To control expression of pathway target genes, the G protein-coupled receptor (GPCR) Smoothened (SMO) activates glioma-associated (GLI) transcription factors via an unknown mechanism. Here we show that, rather than conforming to traditional GPCR signaling paradigms, SMO activates GLI by binding and sequestering protein kinase A (PKA) catalytic subunits at the membrane. This sequestration, triggered by GPCR kinase 2 (GRK2)-mediated phosphorylation of SMO intracellular domains, prevents PKA from phosphorylating soluble substrates, releasing GLI from PKA-mediated inhibition. Our work provides a mechanism directly linking Hh signal transduction at the membrane to GLI transcription in the nucleus. This process is more fundamentally similar between species than prevailing hypotheses suggest. The mechanism described here may apply broadly to other GPCR- and PKA-containing cascades in diverse areas of biology.Competing Interest StatementThe authors have declared no competing interest.View Full Text

biochemistry↗

bioRxiv2017

Limited contribution of rare, noncoding variation to autism spectrum disorder from sequencing of 2,076 genomes in quartet families

Genomic studies to date in autism spectrum disorder (ASD) have largely focused on newly arising mutations that disrupt protein coding sequence and strongly influence risk. We evaluate the contribution of noncoding regulatory variation across the size and frequency spectrum through whole genome sequencing of 519 ASD cases, their unaffected sibling controls, and parents. Cases carry a small excess of de novo (1.02-fold) noncoding variants, which is not significant after correcting for paternal age. Assessing 51,801 regulatory classes, no category is significantly associated with ASD after correction for multiple testing. The strongest signals are observed in coding regions, including structural variation not detected by previous technologies and missense variation. While rare noncoding variation likely contributes to risk in neurodevelopmental disorders, no category of variation has impact equivalent to loss-of-function mutations. Average effect sizes are likely to be smaller than that for coding variation, requiring substantially larger samples to quantify this risk.

genomics↗