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MBC in Press, published online ahead of print February 22, 2006
Mol. Biol. Cell 10.1091/mbc.E05-10-0985

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Submitted on October 26, 2005
Revised on February 13, 2006
Accepted on February 14, 2006

POLKADOTS Are Foci of Functional Interactions in T-Cell Receptor-mediated Signaling to NF-{kappa}B

Jeremy S. Rossman,* Natalia G. Stoicheva,* Felicia D. Langel,* George H. Patterson,{dagger} Jennifer Lippincott-Schwartz,{dagger} and Brian C. Schaefer*

*Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814; {dagger}Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20814

Monitoring Editor: Mark Ginsberg

Stimulation of the T-cell receptor (TCR) results in the activation of several transcription factors, including NF-{kappa}B, that are crucial for T-cell proliferation and gain of effector functions. On TCR engagement, several proteins within the TCR-directed NF-{kappa}B signaling pathway undergo dynamic spatial redistribution, but the significance of these redistribution events is largely unknown. We have previously described TCR-induced cytoplasmic structures called POLKADOTS that are enriched in the NF-{kappa}B signaling intermediate, Bcl10. We now show that these structures are formed only under conditions that promote efficient NF-{kappa}B activation. Furthermore, POLKADOTS formation is dependent upon functional domains of specific NF-{kappa}B signal transducers. Through use of a photoactivatable GFP, we demonstrate that POLKADOTS contain both a highly stable and a rapidly equilibrating protein component. FRET analyses show that POLKADOTS are sites of enriched interactions between Bcl10 and partner signaling proteins. These observations strongly suggest that POLKADOTS are focal sites of dynamic information exchange between cytosolic intermediates in the process of TCR activation of NF-{kappa}B.

Address correspondence to: Brian C. Schaefer (bschaefer{at}usuhs.mil)




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