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PMID: 8628296 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Evidence for posttranscriptional regulation of C/EBPalpha and C/EBPbeta isoform expression during the lipopolysaccharide-mediated acute-phase response.

Molecular and cellular biology ·Vol. 16 ·No. 5 ·1996-05-00 ·Pages 2295-306

An MR, Hsieh CC, Reisner PD, Rabek JP, Scott SG, Kuninger DT, Papaconstantinou J

Abstract

The mRNAs of the CCAAT/enhancer-binding trans-activator proteins (C/EBPalpha and C/EBPbeta) serve as templates for the differential translation of several isoforms which have specific transcriptional regulatory functions. By using an oligonucleotide corresponding to the C/EBP binding site of the mouse alpha1-acid glycoprotein promoter, we detected multiple forms of C/EBPalpha and C/EBP++ beta proteins in the mouse liver that have DNA-binding activity. By using specific antisera, we detected C/EBPalphas with molecular masses of 42, 38, 30, and 20 kDa that have DNA-binding activity. The pool levels of the 42- and 30-kDa isoforms were high in control nuclear extracts and decreased significantly after lipopolysaccharide (LPS) treatment. The binding activity and protein levels of the 20-kDa isoform are low in controls and increase dramatically after LPS treatment. C/EBPbeta isoforms with molecular masses of 35, 20, and 16 kDa were also detected. The 35-kDa pool level did not change whereas the 20-kDa isoform was strongly induced in response to LPS. Western (immunoblot) and Southwestern (DNA-protein) analyses show that p42 C/EBPalpha forms specific complexes with the alpha1-acid glycoprotein oligonucleotide in control nuclear extract and that p20 C/EBP beta forms complexes in LPS-treated liver. Our studies suggest that synthesis of specific C/EBPalpha and C/EBPbeta isoforms occurred in the normal liver in vivo and that LPS mediated a differential initiation and inhibition of translation at specific AUG sites within each mRNA. The qualitative and quantitative changes in C/EBPalpha and C/EBPbeta isoform pool levels suggest that LPS or an LPS-stimulated factor can regulate the selection of AUG start sites for both activation and repression of translation. This regulation appears to involve an LPS-mediated down-regulation of initiation at the first AUG codon of the 42-kDa C/EBPalpha and dramatic translational up-regulation at the fifth AUG codon of the 20-kDa C/EBPalpha and the third AUG codon of the 20-kDa C/EBPbeta. These regulatory events suggest the existence of proteins that may act as translational trans-acting factors.

MeSH Terms
Amino Acid Sequence Animals Base Sequence CCAAT-Enhancer-Binding Proteins Cell Nucleus/metabolism DNA-Binding Proteins/biosynthesis Electrophoresis, Polyacrylamide Gel Gene Expression/drug effects Immune Sera Immunoblotting Kinetics Lipopolysaccharides/pharmacology Liver/metabolism Male Mice Mice, Inbred BALB C Molecular Sequence Data Nuclear Proteins/biosynthesis,isolation & purification,metabolism Oligodeoxyribonucleotides Orosomucoid/biosynthesis,genetics Peptide Fragments/chemical synthesis,immunology Promoter Regions, Genetic Protein Biosynthesis/drug effects RNA, Messenger/metabolism Transcription Factors/biosynthesis Transcription, Genetic
Chemicals
CCAAT-Enhancer-Binding Proteins DNA-Binding Proteins Immune Sera Lipopolysaccharides Nuclear Proteins Oligodeoxyribonucleotides Orosomucoid Peptide Fragments RNA, Messenger Transcription Factors
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
An M R
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, 77555, USA.
Hsieh C C
Reisner P D
Rabek J P
Scott S G
Kuninger D T
Papaconstantinou J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.5.2295
Published
1996-05-00
Pages
2295-306
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231217
Subset
IM
Grants
NIA NIH HHS · P01 AG10514 · United States
External Links
PubMed source DOI: 10.1128/MCB.16.5.2295 PMC Full Text View journal details
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