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Molecular and Cellular Biology, September 2007, p. 6265-6278, Vol. 27, No. 18
0270-7306/07/$08.00+0     doi:10.1128/MCB.00500-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Analysis of Turnover and Translation Regulatory RNA-Binding Protein Expression through Binding to Cognate mRNAs{triangledown}

Rudolf Pullmann Jr., Hyeon Ho Kim, Kotb Abdelmohsen, Ashish Lal, Jennifer L. Martindale, Xiaoling Yang, and Myriam Gorospe*

Laboratory of Cellular and Molecular Biology, National Institute on Aging-Intramural Research Program, National Institutes of Health, Baltimore, Maryland 21228

Received 22 March 2007/ Returned for modification 24 April 2007/ Accepted 25 June 2007

RNA-binding proteins (RBPs) that associate with specific mRNA sequences and function as mRNA turnover and translation regulatory (TTR) RBPs are emerging as pivotal posttranscriptional regulators of gene expression. However, little is known about the mechanisms that govern the expression of TTR-RBPs. Here, we employed human cervical carcinoma HeLa cells to test the hypothesis that TTR-RBP expression is influenced posttranscriptionally by TTR-RBPs themselves. Systematic testing of the TTR-RBPs AUF1, HuR, KSRP, NF90, TIA-1, and TIAR led to three key discoveries. First, each TTR-RBP was found to associate with its cognate mRNA and with several other TTR-RBP-encoding mRNAs, as determined by testing both endogenous and biotinylated transcripts. Second, silencing of individual TTR-RBPs influenced the expression of other TTR-RBPs at the mRNA and/or protein level. Third, further analysis of two specific ribonucleoprotein (RNP) complexes revealed that TIA-1 expression was controlled via HuR-enhanced mRNA stabilization and TIAR-repressed translation. Together, our findings underscore the notion that TTR-RBP expression is controlled, at least in part, at the posttranscriptional level through a complex circuitry of self- and cross-regulatory RNP interactions.

* Corresponding author. Mailing address: Box 12, LCMB, NIA-IRP, NIH, 5600 Nathan Shock Dr., Baltimore, MD 21224. Phone: (410) 558-8443. Fax: (410) 558-8386. E-mail: myriam-gorospe{at}nih.gov

{triangledown} Published ahead of print on 9 July 2007.

Molecular and Cellular Biology, September 2007, p. 6265-6278, Vol. 27, No. 18
0270-7306/07/$08.00+0     doi:10.1128/MCB.00500-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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