14-3-3 interaction with phosphodiesterase 8A sustains PKA signaling and downregulates the MAPK pathway.

The cAMP/PKA and mitogen-activated protein kinase (MAPK) signaling cascade control many cellular processes and are highly regulated for optimal cellular responses upon external stimuli. Phosphodiesterase 8A (PDE8A) is an important regulator that inhibits signaling via cAMP-dependent PKA by hydrolyzing intracellular cAMP pool. Conversely, PDE8A activates the MAPK pathway by protecting CRAF/Raf1 kinase from PKA-mediated inhibitory phosphorylation at Ser259 residue, a binding site of scaffold protein 14-3-3. It still remains enigmatic as to how the cross-talk involving PDE8A regulation influences cAMP/PKA and MAPK signaling pathways. Here, we report that PDE8A interacts with 14-3-3ζ in both yeast and mammalian system, and this interaction is enhanced upon the activation of PKA, which phosphorylates PDE8A's Ser359 residue. Biophysical characterization of phospho-Ser359 peptide with 14-3-3ζ protein further supports their interaction. Strikingly, 14-3-3ζ reduces the catalytic activity of PDE8A, which upregulates the cAMP/PKA pathway while the MAPK pathway is downregulated. Moreover, 14-3-3ζ in complex with PDE8A and cAMP-bound regulatory subunit of PKA, RIα, delays the deactivation of PKA signaling. Our results define 14-3-3ζ as a molecular switch that operates signaling between cAMP/PKA and MAPK by associating with PDE8A.

Impact of traffic on air pollution in a mid-sized urban city during COVID-19 lockdowns.

In this study, we evaluated the changes in air pollutant concentrations around Milwaukee, WI, during and after lockdown due to the COVID-19 pandemic for a period of 126 days. Measurements of particulate matter (PM1, PM2.5, and PM10), NH3, H2S, and O3 + NO2, were made on a 74-km route of arterial and highway roads from April to August 2020 using a Sniffer 4D sensor mounted to a vehicle. Traffic volume during measurement periods were estimated from smartphone-based traffic data. From lockdown (March 24, 2020-June 11, 2020) to post-lockdown (June 12, 2020-August 26, 2020) median traffic volume increased roughly 30-84%, depending upon the road type. In addition, increases in mean concentrations of NH3 (277%), PM (220-307%), and O3 + NO2 (28%) were also observed. For both traffic and air pollutants, abrupt changes in the data were observed mid-June, shortly after lockdown measures were lifted in Milwaukee County. Indeed, traffic was able to explain up to 57% of PM, 47% of NH3, and 42% of O3 + NO2 variance in pollutant concentrations on arterial and highway road segments. Two arterial roads that did not have statistically significant changes in traffic patterns during the lockdown exhibited no statistically significant trends between traffic and air quality parameters. This study demonstrated that COVID-19 lockdowns in Milwaukee, WI, caused significant decreases in traffic, which in turn had a direct impact on air pollutants. It also highlights the need for traffic volume and air quality data at relevant spatial and temporal scales for accurately assessing source apportionment of combustion-based air pollutants, which cannot be captured with typical ground-based sensor systems.