The ACT-America Earth Venture mission conducted five airborne campaigns across four seasons from 2016-2019, to study the transport and fluxes of Greenhouse gases across the eastern United States (US). Unprecedented spatial sampling of atmospheric tracers (CO2, CO, and COS) related to biospheric processes offers opportunities to improve our qualitative and quantitative understanding of seasonal and spatial patterns of biospheric carbon uptake. Here, we examine co-variation of boundary layer enhancements of CO2, CO, and COS across three diverse regions: the crop-dominated Midwest, evergreen-dominated South, and deciduous broadleaf-dominated Northeast. To understand the biogeochemical processes controlling these tracers, we compare the observed co-variation to simulated co-variation resulting from model- and satellite- constrained surface carbon fluxes. We found indication of a common terrestrial biogenic sink of CO2 and COS and secondary production of CO from biogenic sources in summer throughout the eastern US. Stomatal conductance likely drives fluxes through diffusion of CO2 and COS into leaves and emission of biogenic volatile organic compounds into the atmosphere. ACT-America airborne campaigns filled a critical sampling gap in the southern US, providing information about seasonal carbon uptake in southern temperate forests, and demanding a deeper investigation of underlying biological processes and climate sensitivities. Satellite- constrained carbon fluxes capture much of the observed seasonal and spatial variability, but underestimate the magnitude of net CO2 and COS depletion in the Southeast, indicating a stronger than expected net sink in late summer.