Hippocampal-prefrontal interactions are implicated in working memory (WM) and altered in psychiatric conditions with cognitive impairment such as schizophrenia. While coupling between both structures is crucial for WM performance in rodents, evidence from human studies is conflicting and translation of findings is complicated by the use of differing paradigms across species. We therefore used functional magnetic resonance imaging (fMRI) together with a spatial WM paradigm adapted from rodent research to examine hippocampal-prefrontal coupling in humans. A prefrontal-parietal network was functionally connected to hippocampus (HC) during task stages requiring high levels of executive control but not during a matched control condition. The magnitude of coupling in a network comprising HC, bilateral dorsolateral prefrontal cortex (DLPFC) and right supramarginal gyrus explained one fourth of the variability in an independent spatial WM task but was unrelated to visual WM performance. HC-DLPFC coupling may thus represent a systems-level mechanism specific to spatial WM that is conserved across species, suggesting its utility for modeling cognitive dysfunction in translational neuroscience.Neuropsychopharmacology accepted article preview online, 12 January 2015. doi:10.1038/npp.2015.13.