Spatial organization of chromatin at the nuclear lamina is critical for establishing and maintaining cellular identity. Mechanical inputs are emerging as key regulators of nuclear architecture, but whether and how they influence genome-lamina interactions remain unresolved. We modeled aspects of laminopathies via siRNA-mediated lamin A/C (LMNA) knockdown to examine how the nuclear lamina and cytoskeleton contribute to the loss of lamina-associated domain (LAD) organization. Genomics and imaging analyses reveal spatial positioning of LADs with a specific molecular signature are particularly vulnerable to LMNA reduction. Further, a subset of these LADs retain their lamina-association with either concomitant disruption of the Linker of Nucleoskeleton and Cytoskeleton complex or microtubule depolymerization. Conversely, microtubule stabilization phenocopies spatial positioning changes observed following LMNA-knockdown. These data suggest the balance of nuclear lamina and cytoskeletal interactions across the nuclear membrane regulates peripheral chromatin organization. In the context of a compromised nuclear lamina, such as LMNA reduction, the cytoskeleton contributes to the loss of peripheral chromatin organization.