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Upregulation of Alpha-2-Macroglobulin in Replicative Senescence

  • Li Wei Ma
  • Guo Dong Li
  • Tan Jun Tong
Chapter
Part of the Tumor Dormancy and Cellular Quiescence and Senescence book series (DOQU, volume 2)

Abstract

Replicative senescence is a well-established model system for studying the molecular basis of aging. Using high-density complementary deoxyribonucleic acid (cDNA) arrays, we identified alpha-2-macroglobulin (α2M) as a differentially expressed gene in replicative senescence. Our study demonstrated a positive linear-correlation of the mRNA level of α2M with cumulative population doublings (PDL) of human fibroblasts. The levels of α2M increased in senescence cells, but not in quiescent state of cultured fibroblasts, and remained stable in immortal HeLa cells. Moreover, the mRNA level of α2M in leucocytes showed significant difference between newborn and old human body. These results indicate that the up-regulated expression of α2M could be a universal age-related phenotype and the mRNA level of α2M may be used as a biomarker of aging in vitro and in vivo. To further explore the mechanism responsible for the up-regulation of α2M in senescent, We identified a novel transcriptional regulatory element, the α2M transcription enhancement element (ATEE), within the α2M promoter. This element differentially activates α2M expression in senescent versus young fibroblasts. Electrophoretic mobility shift assays revealed abundant complexes in senescent cell nuclear extracts compared with young cell nuclear extracts. The DNase I footprint revealed the protein-binding core sequence through which the protein binds the ATEE. Mutation within ATEE selectively abolished α2M promoter activity in senescent (but not young) cells. These results indicated the ATEE, as a positive transcription regulatory element, contributes to the up-regulation of α2M during replicative senescence.

Keywords

Alpha-2-macroglobulin (α2Μ) Electrophoretic mobility shift assays (EMSA) Functional promoter region determination α2M transcription enhancement element (ATEE) Positive element identification Replicative senescence Reporter gene assay Trans-acting factors 

Notes

Acknowledgements

This work was supported by grants 2013CB530801 from National Basic Research of China and grants 31000609,81170319 from National Natural Science Foundation of China.

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  1. 1.Research Center on Aging, Department of Biochemistry and Molecular BiologyPeking University Health Science CenterBeijingPeople’s Republic of China

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