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© The Rockefeller University Press, 0021-9525/1998//1801 $5.00
The Journal of Cell Biology, Volume 143, Number 7, , 1998 1801-1812

Functional Analysis of Tpr: Identification of Nuclear Pore Complex Association and Nuclear Localization Domains and a Role in mRNA Export

Peter Bangs^*, Brian Burke, Christine Powers, Roger Craig, Aruna Purohit^*, and Stephen Doxsey^*

^* Program in Molecular Medicine and Department of Molecular Cell Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605; and Department of Cell Biology and Anatomy, University of Calgary, Calgary, Alberta, Canada T2N 4N1

Tpr is a 270-kD coiled-coil protein localized to intranuclear filaments of the nuclear pore complex (NPC). The mechanism by which Tpr contributes to the structure and function of the nuclear pore is currently unknown. To gain insight into Tpr function, we expressed the full-length protein and several subdomains in mammalian cell lines and examined their effects on nuclear pore function. Through this analysis, we identified an NH₂-terminal domain that was sufficient for association with the nucleoplasmic aspect of the NPC. In addition, we unexpectedly found that the acidic COOH terminus was efficiently transported into the nuclear interior, an event that was apparently mediated by a putative nuclear localization sequence. Ectopic expression of the full-length Tpr caused a dramatic accumulation of poly(A)⁺ RNA within the nucleus. Similar results were observed with domains that localized to the NPC and the nuclear interior. In contrast, expression of these proteins did not appear to affect nuclear import. These data are consistent with a model in which Tpr is tethered to intranuclear filaments of the NPC by its coiled coil domain leaving the acidic COOH terminus free to interact with soluble transport factors and mediate export of macromolecules from the nucleus.

Key Words: Tpr • nuclear pore • nuclear localization sequence • nuclear export • mRNA

Abbreviations used in this paper: GFP, green fluorescent protein; HA, hemagglutinin; NE, nuclear envelope; NLS, nuclear localization signal; NPC, nuclear pore complex.

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