Adenovirus E1A proteins direct subcellular redistribution of Nek9, A NimA‐related kinase

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Abstract
A monoclonal antibody raised against adenovirus E1A‐associated cellular proteins recognized Nek9, a NimA‐related protein kinase. Subcellular fractionation and immunofluorescence indicated that Nek9 was primarily cytoplasmic with a small portion located in the nucleus whereas E1A was primarily nuclear. Although co‐immunoprecipitation experiments indicated that nuclear Nek9 interacted, directly or indirectly, with E1A, the major effect of E1A was to diminish the amount of Nek9 in the nucleus suggesting that E1A alters the subcellular distribution of Nek9 and that the interaction is transient. A Nek9 deletion mutant lacking a central RCC1‐like domain interacted stably with E1A and accumulated in the nucleus in the presence of E1A, possibly representing an intermediate stage of the normally transient Nek9/E1A interaction. The interaction of Nek9 with E1A was dependent on the N‐terminal sequences of E1A. Attempts to stably overexpress either Nek9 or the kinase‐inactive mutant in various cell lines were unsuccessful; however, the presence of E1A allowed stable overexpression of both proteins. These results suggest that E1A disrupts a nuclear function of Nek9. J. Cell. Physiol. 212: 13–25, 2007.