The striatum, especially its dorsolateral part, plays a major role in motor skill learning and habit formation, but it is still unclear how this contribution might be mediated at the neuronal level. We recorded single neurons in the posterior putamen of two monkeys performing an overlearned sequence of arm reaching movements to examine whether task-related activities are sensitive to manipulations of the serial order of stimulus-target locations. The monkeys' capacity to learn sequential regularities was assessed by comparing arm movement latencies and saccadic ocular reactions when a fixed repeating sequence was replaced with a random sequence. We examined neurons classified as phasically active projection neurons (PANs) and tonically active presumed cholinergic interneurons (TANs). About one-third of the PANs (35/106, 33%) activated during specific parts of a trial displayed modulations of their level of activation when the sequential structure was changed. This differential activity consisted of either decreases or increases in activity without altering the time period during which task-related activations occurred. In addition, half of the TANs (41/80, 51%) changed their responses to task stimuli with the sequence switch, indicating that the response selectivity of TANs reflects the detection of the context that requires adaptation to changes in the serial order of stimulus presentations. Our findings suggest that task-related changes in activity of projection neurons may be an important factor contributing to the production and adjustment of sequential behavior executed in an automatic fashion, whereas putative interneurons may provide a signal for performance monitoring in specific contexts.