The position of the nucleus in the cytoplasm is a highly regulated process and is required for multiple cellular and developmental processes. Defects on different nuclear positioning events are associated with several pathologies such as muscle and nervous system disorders. In this chapter we describe the current knowledge on the mechanism of nuclear positioning. We discuss how the nucleus connects to the cytoskeleton by nesprins and SUN proteins, how this connection is regulated by Samp1, and how this connection is required for proper nuclear positioning. Furthermore, we discuss how nesprins, SUN, and Samp1 form transmembrane actin-associated nuclear (TAN) lines, novel nuclear envelope structures involved in force transduction during nuclear movement. Finally, we describe the recent evidences suggesting a role for the connection between the nucleus and the cytoskeleton in cancer.