Disease relevance of MF(ALPHA)1

• Human macrophage colony-stimulating factor (hM-CSF) cDNA joined to the leader region of the precursor of the yeast mating pheromone alpha-factor (MF alpha L) was expressed at high levels in BmN cells and in silkworm (Bombyx mori) larvae, using recombinant Bombyx mori nuclear polyhedrosis virus, as a vector [1].
• In contrast, the Klebsiella pneumoniae pullulanase gene was placed under the control of the yeast alcohol dehydrogenase gene promoter (ADC1P) and secreted using the yeast mating pheromone alpha-factor secretion signal (MF alpha 1S) [2].

High impact information on MF(ALPHA)1

• The precursors were translated in a yeast lysate from mRNA obtained by in vitro transcription of the MF alpha 1 and SUC2 genes [3].
• A plasmid carrying the MF alpha gene was identified by screening for production of alpha-factor by mat alpha 2 mutants, which fail to secrete alpha-factor because of simultaneous synthesis and degradation of the factor [4].
• The fusion comprised the prepro region of prepro-alpha-factor (MF alpha 1) N-terminal to phosphoribosyl anthranilate isomerase (TRP1) [5].
• We find by RNA blot analysis that provision of MAT alpha 1 protein permits production of STE3 and MF alpha 1 mRNA in a cells but not in a/alpha cells [6].
• In yeast alpha cells, the product encoded by the MAT alpha 1 gene of the mating-type locus is required for transcription of at least two genes, STE3 and MF alpha 1 [6].

Biological context of MF(ALPHA)1

• Pheromone induction did not require ongoing protein synthesis. a-Factor-induced MF alpha 1 expression was quantitated by analysis of an MF alpha 1::SUC2 fusion gene whose product was assayed for invertase activity [7].
• A genomic disruption of the ARGRIII gene not only leads to an argR phenotype, but also prevents cell growth at 37 degrees C. The disrupted strain is sterile especially in an alpha background and transcription of alpha- and a-specific genes (MF alpha 1 and STE2) is strongly reduced [8].
• Using the MF alpha 1 gene, another chromosome besides chr [9].
• An MF alpha 1-SUC2 (alpha-factor-invertase) gene fusion for study of protein localization and gene expression in yeast [10].
• Similar to MF alpha 1, the MF alpha 1-SUC2 fusion is expressed in alpha haploids at levels 65-75 times higher than in a haploids or in a/alpha diploids; also, high-level expression is eliminated in mat alpha 1 mutants but not in mat alpha 2 mutants [10].

Anatomical context of MF(ALPHA)1

• Furthermore, when the secretion of alpha Gal was directed by the invertase (SUC2) SP, the intracellular enzyme localized to the endoplasmic reticulum (ER), whereas use of the MF alpha 1 SP caused the intracellular enzyme to be outer-chain-glycosylated and processed by the KEX2 endoproteinase, implying that it had passed the ER [11].

Associations of MF(ALPHA)1 with chemical compounds

• Chemostat culture confirmed that growth rate does indeed influence MF alpha 1 promoter activity in glucose-grown cells, the activity of this promoter increasing 2- to 2.5-fold as dilution (growth) rates were reduced from maximal values to 0.2 h-1, but then decreasing with the further decreases in dilution rate needed for fully respiratory growth [12].
• Secretion of pectate lyase (PLe) was directed by the signal sequences of the yeast mating pheromone alpha-factor (MF alpha 1S), B. amyloliquefaciens alpha-amylase (AMYS) and Er. chrysanthemi pectate lyase (pelES) [13].
• The glucose oxidase gene (god) from Aspergillus niger was expressed in Hansenula polymorpha using the methanol oxidase promoter and transcription termination region and the MF-alpha leader sequence from Saccharomyces cerevisiae to direct secretion [14].

Regulatory relationships of MF(ALPHA)1

• The gene isolated in this way was shown also to be essential for the KEX2-independent processing of propheromone overproduced from plasmid-borne MF alpha 1 [15].

Other interactions of MF(ALPHA)1

• A novel aspartyl protease allowing KEX2-independent MF alpha propheromone processing in yeast [15].
• Gene disruption, gene replacement, and promoter analysis of the major alpha-factor structural gene (MF alpha 1) were performed to illustrate the utility of the LYS2 gene for the genetic manipulation of yeasts [16].
• Regulation of alpha-factor production in Saccharomyces cerevisiae: a-factor pheromone-induced expression of the MF alpha 1 and STE13 genes [7].
• In addition, transcripts of the MF alpha 1 and STE3 genes, which encode the alpha-factor precursor and the alpha-factor receptor, respectively, are greatly reduced in this mutant [17].
• These results indicate that at least one alpha-factor structural gene product is required for mating in MAT alpha cells, that MF alpha 1 is responsible for the majority of alpha-factor production, and that MF alpha 1 and MF alpha 2 are the only active alpha-factor genes [18].

References