Object In humans, even a single night of partial sleep deprivation (PSD) can have a negative impact on cognition and affective processing, suggesting that sleep pressure represents a basic physiological constraint of brain function. Among the spontaneously fluctuating resting state networks, the default mode network (DMN) and its anticorrelated network (ACN) hold key functions in segregating internally and externally directed awareness. Task fMRI after sleep deprivation has revealed altered activation patterns in both networks. We hypothesized that effects of PSD in these intrinsically coupled networks can be detected by resting state fMRI. Methods We obtained 6-minute echoplanar imaging time series (1.5 Tesla) during eyes-closed, wakeful-resting experiments from 16 healthy volunteers after normal sleep and after PSD. We used independent component and cross-correlation analysis to study functional connectivity (fc), focusing on the DMN and ACN. Results After PSD, focal reductions of auto-correlation strength were detected in the posterior and anterior midline node of the DMN and in the lateral parietal and insular nodes of the ACN. Cross-correlation analysis confirmed reduced cortico-cortical connectivity within and between the DMN and ACN. Conclusions Increased sleep pressure is reflected in reduced fc of main DMN and ACN nodes during rest. Results have implications for understanding perceptual and cognitive changes after sleep deprivation and are relevant to clinical studies on conditions in which increased sleep propensity is present.