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Vol. 14, Issue 6, 2425-2435, June 2003

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RNA Polymerase III Transcripts and the PTB Protein Are Essential for the Integrity of the Perinucleolar Compartment

Chen Wang *, Joan C. Politz {dagger}, Thoru Pederson {dagger}, and Sui Huang * {ddagger}

* Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611; {dagger} Department of Biochemistry and Molecular Pharmacology and Program in Cell Dynamics, University of Massachusetts Medical School, Worcester, Massachusetts 01605

Submitted December 12, 2002; Accepted January 30, 2003
Monitoring Editor: Joseph Gall

The perinucleolar compartment (PNC) is a nuclear substructure present in transformed cells. The PNC is defined by high concentrations of certain RNA binding proteins and a subset of small RNAs transcribed by RNA polymerase III (pol III), including the signal recognition particle RNA and an Alu RNA as reported here. To determine if the PNC is dependent on pol III transcription, HeLa cells were microinjected with the selective pol III inhibitor, Tagetin. This resulted in disassembly of the PNC, whereas inhibition of pol I by cycloheximide or pol II by {alpha}-amanitin did not significantly affect the PNC. However, overexpression of one of the PNC-associated RNAs from a pol II promoter followed by injection of Tagetin blocked the Tagetin-induced PNC disassembly, demonstrating that it is the RNA rather than pol III activity that is important for the PNC integrity. To elucidate the role of the PNC-associated protein PTB, its synthesis was inhibited by siRNA. This resulted in a reduction of the number of PNC-containing cells and the PNC size. Together, these findings suggest, as a working model, that PNCs may be involved in the metabolism of specific pol III transcripts in the transformed state and that PTB is one of the key elements mediating this process.

Abbreviations used: CMV, cytomegalovirus; CUG-BP, CUG-binding protein; PNC, perinucleolar compartment; PTB, polypyrimidine tract binding; SRP, signal recognition particle; UBF, upstream binding factor.

{ddagger} Corresponding author. E-mail address: s-huang2{at}northwestern.edu.




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