Wrapping DNA into a nucleosome influences factor binding to cognate sites, and thus the positions of nucleosomes in eukaryotic genomes contribute to gene regulation. Nucleosome positioning is influenced by DNA sequence, chromatin remodelers and non-histone chromatin factors, and genomic maps of nucleosomes are now being constructed. However, interpretation of these maps requires consideration of chromatin dynamics, as even some positioned nucleosomes appear subject to rapid unwinding and eviction. The dynamic properties of nucleosomes contribute to several processes, including gene regulation, mechanisms of transcription and the inheritance of chromatin states. Understanding the positions and dynamic behavior of nucleosomes promises to shed light on why transcription factors bind so many fewer sites than predicted, how histone variants may be targeted, and how chromatin states are delineated.