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Journal Article | Research Support, U.S. Gov't, P.H.S.

Transcription-independent turnover of I kappa B alpha during monocyte adherence: implications for a translational component regulating I kappa B alpha/MAD-3 mRNA levels.

A K Lofquist, K Mondal, J S Morris, J S Haskill
A K Lofquist
UNC Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill 27599-7295.
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K Mondal
UNC Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill 27599-7295.
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J S Morris
UNC Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill 27599-7295.
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J S Haskill
UNC Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill 27599-7295.
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DOI: 10.1128/MCB.15.3.1737
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ABSTRACT

We identified I kappa B alpha/MAD-3 as an immediate-early gene in human monocytes that is expressed in response to a variety of signals, including adhesion, lipopolysaccharide, and phorbol myristate acetate. Within 5 min of monocyte adhesion, the level of the I kappa B alpha protein is markedly diminished but is rapidly replaced in a cycloheximide-sensitive manner within 20 min. Accompanying the rapid turnover of the I kappa B alpha protein is simultaneous translocation of NF-kappa B-related transcription factors to nuclei of adhered monocytes. The demonstration that NF-kappa B can regulate I kappa B alpha/MAD-3 gene transcription in other cell types suggested that the rapid increase in steady-state I kappa B alpha/MAD-3 mRNA levels we observed within 30 min of monocyte adherence would result from NF-kappa B-dependent transcriptional stimulation of the I kappa B alpha/MAD-3 gene. Nuclear run-on analyses indicated that, instead, while several immediate-early cytokine genes, such as the interleukin 1 beta (IL-1 beta) gene, were transcriptionally activated during monocyte adhesion, the rate of I kappa B alpha/MAD-3 gene transcription remained constant. The adherence-dependent increase in I kappa B alpha/MAD-3 mRNA levels was also not a consequence of mRNA stabilization events. Interestingly, while increases in both IL-1 beta and I kappa B alpha/MAD-3 mRNA levels were detected in nuclei of adherent monocytes, cytoplasmic levels of IL-1 beta mRNA increased during adherence whereas those of I kappa B alpha/MAD-3 mRNA did not. Taken together, our data suggest that two interactive mechanisms regulate monocytic I kappa B alpha/MAD-3 mRNA levels. We propose that adherent monocytes regulate nuclear processing (or decay) of I kappa B alpha/MAD-3 mRNA, thereby increasing mRNA levels without stimulating I kappa B alpha/MAD-3 gene transcription. Moreover, since inhibition of protein synthesis leads to accumulation of I kappa B alpha/MAD-3 mRNA without stimulating I kappa B alpha/MAD-3 gene transcription, we suggest that low cytoplasmic levels of I kappa B alpha/MAD-3 mRNA are maintained by a translation-dependent degradation mechanism.

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Transcription-independent turnover of I kappa B alpha during monocyte adherence: implications for a translational component regulating I kappa B alpha/MAD-3 mRNA levels.
A K Lofquist, K Mondal, J S Morris, J S Haskill
Molecular and Cellular Biology Mar 1995, 15 (3) 1737-1746; DOI: 10.1128/MCB.15.3.1737

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Transcription-independent turnover of I kappa B alpha during monocyte adherence: implications for a translational component regulating I kappa B alpha/MAD-3 mRNA levels.
A K Lofquist, K Mondal, J S Morris, J S Haskill
Molecular and Cellular Biology Mar 1995, 15 (3) 1737-1746; DOI: 10.1128/MCB.15.3.1737
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