Ultraviolet radiation increases tropoelastin accumulation by a post-transcriptional mechanism in dermal fibroblasts

Chronically sun-damaged human skin is characterized by dermal connective tissue damage that includes the massive accumulation of abnormal elastic fibers. The content of elastin, the major protein component of elastic fibers, is increased two- to sixfold in sun-damaged skin. The aim of this study was to determine the mechanism responsible for the increase in elastin levels after ultraviolet (UV) irradiation. Confluent cultures of normal dermal fibroblasts were irradiated with 4.5 mJ/cm2 of UVB; sham-treated cells served as the control group. The accumulation of tropoelastin was determined at 5 d after treatment by measuring the incorporation of 14C-proline into radiolabeled tropoelastin isolated from cell layers and media. UV irradiation increased radiolabeled tropoelastin accumulation approximately twofold without affecting DNA content, the total amount of radiolabeled protein, or tropoelastin secretion. Moreover, the steady-state levels of tropoelastin mRNA, as determined by slot blot hybridizations, were unaffected by UV treatment. However, the translation of tropoelastin mRNA was increased when total RNA from irradiated cells was used in cell-free translation experiments. These results suggest that altered translational efficiency may account for the increase in tropoelastin accumulation after UV irradiation. In support of this hypothesis, nucleotide sequences were derived from tropoelastin mRNA isolated from UV-irradiated and nonirradiated dermal fibroblasts. Almost a 12% substitution rate was observed in nucleotide sequences derived from the 3' untranslated region of tropoelastin mRNA from the UV-treated cells. In contrast, a coding domain of tropoelastin did not contain base-substitution mutations. These multiple base substitutions in a noncoding domain of tropoelastin mRNA may be responsible for the post-transcriptional increase in tropoelastin accumulation after UV irradiation.