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(Received for publication, April 11, 1997)
From the Division of Pulmonary and Critical Care Medicine,
Departments of Medicine and Pathology, University of Maryland School of
Medicine, Baltimore, Maryland 21201, Medical Service, Baltimore
Veterans Administration Medical Center, Baltimore, Maryland 21201, and
the University of Maryland at Baltimore Cytokine Core Laboratory,
Baltimore, Maryland 21201
In unstimulated RAW 264.7 macrophage-like cells,
tumor necrosis factor-
Volume 272, Number 34,
Issue of August 22, 1997
pp. 21120-21127
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
Poly(A) Tail Metabolism in Tumor Necrosis
Factor-
Regulation
(TNF-) mRNA was transcribed and
accumulated in the cytoplasm, but the TNF- transcripts failed to
associate with polysomes, and TNF- protein was not detected.
Stimulation with lipopolysaccharide (LPS) induced an increase in
TNF- transcription, cytoplasmic TNF- mRNA accumulation,
polysome association, and secretion of TNF- protein. This process
was associated with a 200-nucleotide increase in the apparent length of
the TNF- mRNA. The difference in TNF- mRNA size was
caused by marked truncation of the 3
poly(A) tail in unstimulated
cells. Fully adenylated TNF- mRNA appeared within 15 min of
LPS stimulation. We speculate that removal of the poly(A) tail blocks
initiation of TNF- translation in unstimulated macrophages. LPS
inactivates this process, allowing synthesis of translatable
polyadenylated TNF- mRNA.
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