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Related Experiment Videos

Defining the regulatory factors required for epidermal gene expression.

S Sinha1, L Degenstein, C Copenhaver

  • 1Howard Hughes Medical Institute, Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois 60637, USA.

Molecular and Cellular Biology
|March 14, 2000
PubMed
Summary

Keratin 14 (K14) gene regulation in epidermis was studied using transgenic mice and cell cultures. Key regulatory elements were identified within the K14 gene

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Area of Science:

  • Molecular Biology
  • Genetics
  • Dermatology

Background:

  • Keratins K5 and K14 are crucial for stratified epithelia.
  • Their high-level, tissue-specific transcription offers insight into epidermis regulation.

Purpose of the Study:

  • To identify regulatory mechanisms of epidermis-specific transcription for the K14 gene.
  • To elucidate the role of regulatory elements in keratinocyte gene expression.

Main Methods:

  • Analysis of DNase I-hypersensitive sites (HSs) in K14 5' regulatory sequences.
  • In vivo transgenic approaches and in vitro keratinocyte culture.
  • Electrophoretic mobility shift and mutational assays.

Main Results:

  • Identified a 700-bp regulatory domain containing HSsII and -III critical for K14 expression.

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  • A 125-bp fragment with HSsII demonstrated enhancer activity.
  • AP-1, ets, and AP-2 transcription factors are involved in K14 regulation within HSsII.

    • Conclusions:

    • A novel enhancer region regulates K14 transcription in keratinocytes.
    • Specific transcription factors (ets, AP-2) within HSsII are key for epidermal specificity.
    • Further investigation is needed to fully restrict activity to keratinocytes in vivo.