UC San Diego

The nucleus is in many ways the centerpiece of the eukaryotic cell, as it houses the genome and is the primary site of gene regulation. Nuclear enclosure is achieved by the double lipid bilayer named the nuclear envelope (NE). The outer membrane of the NE is connected and continuous with the endoplasmic reticulum (ER). The inner membrane of the NE attaches to chromatin and a meshwork of intermediate filaments called the nuclear lamina though NE-specific integral membrane proteins. Transport between the cytoplasm and nucleoplasm is mediated by nuclear pore complexes, multi-protein assemblies that are present where of the NE where the outer and inner membranes are connected. In metazoans, the nuclear envelope is broken down during mitosis to allow for cytoplasm spindle formation and segregation the NE materials into the daughter cells. At the beginning of my thesis the fate of NE components during cell division and the mechanism of nuclear reformation have been controversial, and it was unclear whether the NE is broken into vesicles or absorbed into the ER during mitosis. The main focus of this thesis was to characterize NE formation at the end of open mitosis. We determine that the network of ER tubules directly contributes to nuclear membrane formation in a fusion independent mechanism. A role for the ER shaping protein family of Reticulons as a negative regulator of NE formation was also characterized. A model was developed where transmembrane proteins of the NE target and reshape ER membranes around chromatin during nuclear assembly. This model was supported be a detailed kinetic analysis of nuclear assembly in cells where protein expression levels of candidate proteins were changed. Together these studies clarify the mechanism of how the nuclear membrane encloses the chromatin mass at the end of cell division