解脂耶氏酵母中囊泡蛋白YlSec15的鉴定及功能研究 <sup>*</sup>

doi:10.13523/j.cb.20190305

中国生物工程杂志

2019, Vol. 39

Issue (3): 29-36 DOI: 10.13523/j.cb.20190305

研究报告

解脂耶氏酵母中囊泡蛋白YlSec15的鉴定及功能研究 ^*

陈凯丽¹,张付涛¹,王东月¹,张倩¹,李运清^2,^**(

)

1 济宁医学院临床医学院济宁 272067
2 济宁医学院病原生物学教研室济宁 272067

Identification and Characterization of the Vesicle Protein YlSec15 in Yarrowia lipolytica

Kai-li CHEN¹,Fu-tao ZHANG¹,Dong-yue WANG¹,Qian ZHANG¹,Yun-qing LI^2,^**(

)

1 School of Clinical Medicine, Jining Medical University, Jining 272067,China
2 Department of Pathogenic Biology, Jining Medical University, Jining 272067, China

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摘要：

解脂耶氏酵母(Yarrowia lipolytica)进行出芽繁殖时,决定未来分裂平面的出芽位点不是随机选取的,而是选择在前一次细胞分裂位置的对侧出芽,即进行双极出芽。目前对解脂耶氏酵母双极出芽的分子调控机制并不清楚。通过观察蛋白定位及过量表达的方法研究了解脂耶氏酵母中囊泡蛋白YlSec15的功能。结果表明:YlSec15在细胞中有明显的极性定位,在细胞的小芽内以及大中芽的芽颈处富集,过量表达YlSec15抑制了菌丝的形成并使得部分细胞的出芽位点选择方式由双极出芽转变为随机出芽,而引起这一变化的原因可能是由于过量的YlSec15在细胞中不能进行正常的极性定位。此外,YlSec15可能是通过YlRas2介导的信号通路参与调控细胞的菌丝形成及双极出芽。这一发现丰富了解脂耶氏酵母中双极出芽的分子调控机制,也证明了极性生长与囊泡运输之间是相互影响的。

关键词： 解脂耶氏酵母; 双极出芽; 菌丝; 囊泡蛋白; YlSec15

Abstract:

Yarrowia lipolytica is a budding yeast and the budding position which determined the division plane is not selected randomly but determined by the distance of the last division site, that is, bud in the bipolar budding pattern. At present, the molecular regulation pathway of the bipolar budding is not well studied. In this report, the characterization of the vesicle protein YlSec15 was studied through the localization and overexpression. YlSec15 can locate inside the small buds and the neck of the middle or big buds, indicating the patterns of polar localization. Overexpression of YlSec15 in the yeast can inhibit the formation of hyphae and change budding pattern of cells, from bipolar to random. The causes of these changes may be due to the delocalization to the polar site of the excessive YlSec15. Besides, the normal localization of YlSec15 dependent on Ylras2. Based on this result, we speculate that YlSec15 may be involved in regulating the polar growth of cells through the signal pathway which is mediated by YlRas2. The discovery in this paper will enrich the molecular regulation mechanism of the bipolar budding and confirm the relationship between polar growth and vesicle transport in Yarrowia lipolytica.

Key words: Yarrowia lipolytica Bipolar budding Hyphae Vesicle protein YlSec15

收稿日期: 2018-09-27 出版日期: 2019-04-12

ZTFLH:

Q71

基金资助: * 国家自然科学基金青年基金资助项目(31500056)

通讯作者: 李运清 E-mail: liyunqing2013@whu.edu.cn

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引用本文:

陈凯丽,张付涛,王东月,张倩,李运清. 解脂耶氏酵母中囊泡蛋白YlSec15的鉴定及功能研究 ^*[J]. 中国生物工程杂志, 2019, 39(3): 29-36.

Kai-li CHEN,Fu-tao ZHANG,Dong-yue WANG,Qian ZHANG,Yun-qing LI. Identification and Characterization of the Vesicle Protein YlSec15 in Yarrowia lipolytica. China Biotechnology, 2019, 39(3): 29-36.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20190305 或 https://manu60.magtech.com.cn/biotech/CN/Y2019/V39/I3/29

菌株	基因型	来源
PO1a	MATA leu2-270 ura3-302	13
YLX260	MATA leu2-270 ura3-302 Ylrsr1∷loxR/P	8
YLX81	MATA leu2-270 ura3-302 Ylras2	9
YLJ4	MATA leu2-270 ura3-302 [Integrative URA3 YlSec15↑]	This study
YLJ8	MATA leu2-270 ura3-302 [CEN LEU2 YlSEC15-EGFP]	This study
YLJ9	MATA leu2-270 ura3-302 [CEN LEU2 EGFP-YlSec15↑]	This study
DH5	F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15Δ (lacZYA-argF) U169, hsdR17 ( $r K ?$ $m K +$ ), λ-	Takara, Japan

表1 实验中所用的菌株

表2 实验中所用的主要引物

图1 YlSec15的分离及鉴定

图2 YlSec15-EGFP在细胞中的定位

图3 YlSec15参与调控细胞的形态

图4 过量表达YlSec15影响细胞的双极出芽

图5 过量的YlSec15在细胞中的分布

图6 YlSec15-EGFP在Ylrsr1Δ和Ylras2Δ中的定位

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