Part:BBa_K5143009

Alpha-Factor secretion signal V1, optimized for Saccharomyces cerevisiae

Description

The α-mating factor is a peptide hormone produced by the yeast Saccharomyces cerevisiae, which is crucial for the mating process of these yeast cells. Here are the key details about the α-mating factor:

• Function: It is involved in the mating process of S. cerevisiae by being secreted by α-type yeast cells to attract a-type cells, facilitating the mating interaction.
• Production: Synthesized as a pre-protein that includes a signal peptide and the mature α-mating factor peptide.
• Structure and Processing:
  • Pre-Pro-α-Mating Factor: Initially synthesized with a signal peptide (pre-region) that directs the nascent protein to the secretory pathway.
  • Mature Peptide: After cleavage of the signal peptide, the remaining peptide is processed into the active α-mating factor.
  • Processing: Includes cleavage of the signal peptide and pro-region to yield the mature peptide.
• Secretion:
  • Signal Peptide Function: Directs the protein to the endoplasmic reticulum (ER), where it is cleaved for passage through the secretory pathway.
  • Mature Peptide Secretion: Transported through the Golgi apparatus and secreted into the extracellular environment.
• Applications in Biotechnology:
  • Protein Secretion: The α-mating factor's signal peptide is used to enhance the secretion of various proteins in yeast by attaching it to a protein of interest.
  • Engineering Yeast Strains: Yeast strains can be engineered for efficient production of heterologous proteins by incorporating the α-mating factor signal peptide into their expression constructs.

Construction

The Alpha-Factor gene was synthesised and its nucleotide sequence optimised for DNA synthesis and expression in Saccharomyces cerevisiae. The gene exists in 2 versions: which limits homologous recombination in the Saccharomyces cerevisiae genome. The second version can be found here : BBa_K5143010
These two versions of the alpha-factor can be found in the following composite part: BBa_K5143023

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1361
  Illegal PstI site found at 2789
  Illegal PstI site found at 3092
  Illegal PstI site found at 3185
  Illegal PstI site found at 3191
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1361
  Illegal PstI site found at 2789
  Illegal PstI site found at 3092
  Illegal PstI site found at 3185
  Illegal PstI site found at 3191
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1361
  Illegal BamHI site found at 1906
  Illegal BamHI site found at 2620
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1361
  Illegal PstI site found at 2789
  Illegal PstI site found at 3092
  Illegal PstI site found at 3185
  Illegal PstI site found at 3191
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1361
  Illegal PstI site found at 2789
  Illegal PstI site found at 3092
  Illegal PstI site found at 3185
  Illegal PstI site found at 3191
  Illegal NgoMIV site found at 1876
  Illegal AgeI site found at 1915
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

Brake, A. J. et al. Alpha-factor-directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences 81, 4642–4646 (1984).

Aza, P., Molpeceres, G., de Salas, F. & Camarero, S. Design of an improved universal signal peptide based on the α-factor mating secretion signal for enzyme production in yeast. Cell Mol Life Sci 78, 3691–3707 (2021).