The genetic variability and covalent modifications associated with the amino terminus of the protein kinase A (PKA) catalytic (C) subunit suggest that it may contribute to protein-protein interactions and͞or localization. By using a yeast two-hybrid screen, we identified a PKA-interacting protein (AKIP1) that binds to the amino terminus (residues 1-39) of the C subunit of PKA. The interaction was localized to the A helix (residues 14 -39) of the C subunit and to the carboxyl terminus of AKIP1. AKIP1 thus defines the amino-terminal A helix of PKA as a protein interaction motif. In normal breast (Hs 578 Bst) and HeLa cells, AKIP1 is present in the nucleus as speckles. A nuclear localization signal (Arg-14 and Arg-15) was identified. Upon stimulation with forskolin, HeLa cells expressing AKIP1 accumulated higher levels of the endogenous C subunit in the nucleus. Deletion of the carboxyl terminus of AKIP1 or overexpression of residues 1-39 of the C subunit abolished nuclear localization of the activated endogenous C subunit. Thus, AKIP1 describes a PKA-interacting protein that can contribute to localization by a mechanism that is distinct from A-kinase anchoring proteins that interact with the regulatory subunits.nuclear retention ͉ speckles

We demonstrate high-resolution, high-speed 3D nanoparticle tracking using angled micromirrors. When angled micromirrors are introduced into the field of view of an optical microscope, reflected side-on views of a diffusing nanoparticle are projected alongside the usual direct image. The experimental design allows us to find the 3D particle trajectory using fast, centroid-based image processing, with no nonlinear computing operations. We have tracked polystyrene particles of 190 nm diameter with position measurement precision <20 nm in 3D with 3 ms frame duration (i.e., at an imaging rate >330 frames per second). Because the image processing requires only approximately 1 ms per frame, this technique could enable real-time feedback-controlled nanoparticle assembly applications with nanometer precision.

We demonstrate a method using photoactivation localization microscopy (PALM) in a soft-material system, with a rhodamine-lactam dye that is activated by both ultraviolet light and protonation, to reveal the nanoscale photoacid distribution in a model photoresist. Chemically amplified resists are the principal lithographic materials used in the semiconductor industry. The photoacid distribution generated upon exposure and its subsequent evolution during post-exposure bake is a major limiting factor in determining the resolution and lithographic quality of the final developed resist image. Our PALM data sets resolve the acid distribution in a latent image with subdiffraction limit accuracy. Our overall accuracy is currently limited by residual mechanical drift.

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