This study analyzed the effects of expressing human gene 4-hydroxyphenylpyruvate dioxygenase (HPD) on sporulation and spore wall assembly in Saccharomyces cerevisiae. HPD was introduced into S. cerevisiae, and the phenotype differences between humanized HPD yeast and wild-type yeast were compared. It was found that compared with wild-type yeast, humanized HPD yeast had no effect on the sporulation rate, while the fluorescence intensity of spores decreased, ether sensitivity increased, and the fluorescence intensity of calcofluor white increased. This indicated that HPD caused the defect of spore wall and part of the dityrosine layer missing during the assembly process. Moreover, expressed HPD gene changed the sporulation medium color and increased the fluorescence intensity, as homogentisate, the catalytic product of HPD, was oxidized during sporulation. Furthermore, a large number of dityrosine complex leaked into the sporulating medium and dityrosine layer could not assemble correctly in consequence. Overall, this study provides a new insight into the mechanisms of forming dityrosine layer in S. cerevisiae.
WANG Xiaowen
,
YIN Zheng
,
GAO Xiaodong
,
NAKANISHI Hideki
. Expression of human HPD gene interferes correct assembly of the dityrosine layer on the spore wall of Saccharomyces cerevisiae[J]. Food and Fermentation Industries, 2019
, 45(15)
: 17
-23
.
DOI: 10.13995/j.cnki.11-1802/ts.020633
[1] BRIZA P, BREITENBACH M, ELLINGER A, et al. Isolation of two developmentally regulated genes involved in spore wall maturation in Saccharomyces cerevisiae[J]. Genes Dev,1990, 4(10):1 775-1 789.
[2] 王彩虹,高晓冬,中西秀树.筛选抑制酿酒酵母生长的人类基因[J]. 微生物学报,2018, 58(11):10-22.
[3] 殷政,中西秀树,李子杰,等. 利用酿酒酵母筛选特异性抑制产孢的人类基因[J]. 食品与生物技术学报,2017.
[4] 殷政. 筛选特异性抑制酿酒酵母产孢的人类基因[D]. 无锡:江南大学,2017.
[5] FELDER T, BOGENGRUBER E, TENREIRO S, et al. Dtrlp, a multidrug resistance transporter of the major facilitator superfamily, plays an essential role in spore wall maturation in Saccharomyces cerevisiae[J]. Eukaryotic Cell,2002, 1(5):799-810.
[6] MORAN G R. 4-Hydroxyphenylpyruvate dioxygenase[J]. Arch Biochem Biophys,2005, 433(1):117-128.
[7] PHORNPHUTKUL C, INTRONE W J, PERRY M B, et al. Natural history of alkaptonuria[J]. N Engl J Med, 2002, 347(26):2 111-2 121.
[8] MITCHELL A P. Control of meiotic gene expression in Saccharomyces cerevisiae[J]. Microbiological Reviews,1994, 58(1):56-70.
[9] BRIZA P, ECKERSTORFER M, BREITENBACH M. The sporulation-specific enzymes encoded by the DIT1 and DIT2 genes catalyze a two-step reaction leading to a soluble LL-dityrosine-containing precursor of the yeast spore wall[J]. Proc Natl Acad Sci U S A,1994, 91(10):4 524-4 528.
[10] JAMIESON D J. A course on yeast genetics[J]. Trends in Biotechnology,1991, 9(1):364.
[11] NEIMAN A M. Ascospore formation in the yeast Saccharomyces cerevisiae[J]. Microbiol Mol Biol Rev,2005, 69(4):565-584.
[12] ZHANG H, TACHIKAWA H, GAO X D, et al. Applied usage of yeast spores as chitosan beads[J]. Appl Environ. Microbiol, 2014, 80(16):5 098-5 105.
[13] SHI L B, LI Z J, TACHIKAWA H, et al. Use of yeast spores for microncapsulation of enzyme[J]. Appl Environ Microbiol, 2014,80(15):4 502-4 510.
[14] LIN C P, KIM C, SMITH S O, et al. A highly redundant gene network controls assembly of the outer spore wall in S. cerevisiae[J]. Plos Genet,2013, 9(8):e1003700.
[15] BEMENA L D, MUKAMA O, NEIMAN A M, et al. In vitro reconstitution of the yeast spore wall dityrosine layer discloses the mechanism of its assembly[J]. Journal of Biological Chemistry, 2017, 292(38):15 880-15 891.
[16] SHI L B, LI Z J, TACHIKAWA H, et al. Use of yeast spores for microencapsulation of enzymes[J]. Applied and Environmental Microbiology, 2014, 80(15):4 502-4 510.
[17] PAN H P, WANG N, TACHIKAWA H, et al. Osw2 is required for proper assembly of glucan and/or mannan layers of the yeast spore wall[J]. The Journal of Biochemistry,2018, 163(4):293-304.
[18] PAN H P, WANG N, TACHIKAWA H, et al. beta-1,6-glucan synthesis-associated genes are required for proper spore wall formation in Saccharomyces cerevisiae[J]. Yeast, 2017, 34(11):431-446.
