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December 1, 1980Proceedings of the National Academy of SciencesOpen Access

Two interferon mRNAs in human fibroblasts: in vitro translation and Escherichia coli cloning studies.

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Authors

JWJ. WeissenbachLeiden UniversityYCYuti ChernajovskyUniversity of LondonMZM. ZeeviTel Aviv University

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Implication

Laboratory study reveals two distinct non-cross-hybridizing beta-interferon mRNAs in human fibroblasts, indicating divergent molecular regulation and coding sequences.

Key Points

  • Characterize, translate, and clone distinct messenger RNA species that encode biologically active interferon in human fibroblasts.
  • Isolated mRNA from human fibroblasts and separated species using size fractionation.
  • Cloned fractionated mRNA into Escherichia coli plasmid pBR322 and characterized translation products using cell-free protein synthesis systems.
  • Evaluated mRNA induction and sequence homology using cross-hybridization assays following stimulation with poly(rI.rC) or cycloheximide.
  • Hu IFN-beta 1 was identified as an 11S mRNA of 900 nucleotides encoding a 20,000 Mr protein, while Hu IFN-beta 2 was identified as a 14S mRNA of approximately 1,300 nucleotides encoding a 23,000-26,000 Mr protein.
  • Nucleic acid hybridization demonstrated that Hu IFN-beta 1 and Hu IFN-beta 2 mRNAs do not cross-hybridize.
  • Both mRNAs were induced by poly(rI.rC), but cycloheximide alone induced IFN-beta 2 mRNA to approximately 10% of maximum levels without inducing IFN-beta 1.

Cite This Study

Weissenbach et al. (1980) studied this question.

synapsesocial.com/papers/6a0daefbcae7912d2fa53211https://doi.org/10.1073/pnas.77.12.7152
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