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A cDNA encoding a human CCAAT-binding protein cloned by functional complementation in yeast.

D M Becker1, J D Fikes, L Guarente

  • 1Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.

Proceedings of the National Academy of Sciences of the United States of America
|March 1, 1991
PubMed
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Researchers cloned the human Hap2 transcription factor using yeast complementation. This human protein, involved in gene regulation, shows potential for both general and cell-specific functions.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Gene expression regulation is crucial for cellular function.
  • Transcription factors play a key role in controlling gene expression.
  • The yeast transcription factor Hap2 (HAP2) is essential for cellular processes.

Purpose of the Study:

  • To clone the human homolog of the yeast CCAAT-binding transcription factor, Hap2.
  • To characterize the human Hap2 homolog and its role in gene expression.

Main Methods:

  • Construction of a HeLa cell cDNA library expressed in Saccharomyces cerevisiae.
  • Functional complementation of a yeast hap2 mutation to identify the human homolog.
  • Sequence analysis of the cloned human cDNA and its encoded protein.

Related Experiment Videos

  • Analysis of human Hap2 transcript expression in various cell types.

    • Main Results:

    • Successfully cloned the human Hap2 homolog using functional complementation in yeast.
    • The human Hap2 homolog encodes a 257-amino acid protein with a conserved C-terminal region and a glutamine-rich N-terminus.
    • Human Hap2 exhibits distinct transcript patterns, including a ubiquitous 4.1-kb species, a B lymphocyte-specific 7.0-kb species, and a HeLa cell-preferential 1.6-kb species.
    • The glutamine-rich N-terminus suggests a role in transcriptional activation.

    Conclusions:

    • The human Hap2 homolog is a functional transcription factor with potential roles in both constitutive and cell-type-specific gene regulation.
    • The cloning strategy by complementation is effective for isolating human genes with known yeast counterparts.
    • Further research into human Hap2 and related factors could elucidate complex gene regulatory networks.