Regulation of type I collagen mRNA in lung fibroblasts by cystine availability.
The steady-state level of alpha1(I) collagen mRNA is regulated by amino acid availability in human lung fibroblasts. Depletion of amino acids decreases alpha1(I) collagen mRNA levels and repletion of amino acids induces rapid re-expression of alpha1(I) mRNA. In these studies, we examined the requirements for individual amino acids on the regulation of alpha1(I) collagen mRNA. We found that re-expression of alpha1(I) collagen mRNA was critically dependent on cystine but not on other amino acids. However, the addition of cystine alone did not result in re-expression of alpha1(I) collagen mRNA. Following amino acid depletion, the addition of cystine with selective amino acids increased alpha1(I) collagen mRNA levels. The combination of glutamine and cystine increased alpha1(I) collagen mRNA levels 6.3-fold. Methionine or a branch-chain amino acid (leucine, isoleucine or valine) also acted in combination with cystine to increase alpha1(I) collagen mRNA expression, whereas other amino acids were not effective. The prolonged absence of cystine lowered steady-state levels of alpha1(I) collagen mRNA through a mechanism involving decreases in both the rate of gene transcription as assessed by nuclear run-on experiments and mRNA stability as assessed by half-life determination in the presence of actinomycin D. The effect of cystine was not mediated via alterations in the level of glutathione, the major redox buffer in cells, as determined by the addition of buthionine sulphoximine, an inhibitor of gamma-glutamylcysteine synthetase. These data suggest that cystine directly affects the regulation of alpha1(I) collagen mRNA.
Rishikof
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DC
Pulmonary Center, Boston University School of Medicine, 80 East Concord Street, Boston, MA 02118, USA.
Kuang
P P
PP
Poliks
C
C
Goldstein
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RH
D000597:Amino Acids, Branched-Chain / Q000008:administration & dosage; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics*; D003470:Culture Media; D003553:Cystine / Q000008:administration & dosage*; D004355:Drug Stability; D004622:Embryo, Mammalian; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D005721:Glutamate-Cysteine Ligase / Q000037:antagonists & inhibitors; D005973:Glutamine / Q000008:administration & dosage; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000378:metabolism*; D008715:Methionine / Q000008:administration & dosage; D010084:Oxidation-Reduction; D012333:RNA, Messenger / Q000378:metabolism*
D000597:Amino Acids, Branched-Chain; D003470:Culture Media; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D005973:Glutamine; D003553:Cystine; D019328:Buthionine Sulfoximine; D003094:Collagen; D008715:Methionine; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione