Analysis of Martrix Targeting Sequence of Human Mitochondrial OGT in <i>Saccharomyces cerevisiae</i>

doi:10.13523/j.cb.20190405

China Biotechnology

2019, Vol. 39

Issue (4): 32-37 DOI: 10.13523/j.cb.20190405

Analysis of Martrix Targeting Sequence of Human Mitochondrial OGT in Saccharomyces cerevisiae

Feng LI,Xiao-dong GAO,Hideki NAKANISHI(

)

Key Laboratory of Carbohydrate Chemistry and Biotechnology, Jiangnan University, Wuxi 214122, China

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Abstract

Human mitochondrial O-GlcNAc transferase (mOGT) localized in mitochondria inner membrane through its N-terminal martrix targeting sequence(MTS), mOGT overexpression caused cell spoptosis. O-GlcNAc modification existed in most eukaryote cells except Saccharomyces cerevisiae. In order to analyze mOGT induced mammalian cell apoptosis, the mechanism of mOGT caused yeast cell growth defect was investigated. Herein, mOGT was overexpressed in Saccharomyces cerevisiae. Both yeast growth defect and mOGT localization were largely depends on MTS sequence. Furthermore, MTS expression caused yeast mitochondria fussion. Therefore, MTS overexpressed yeast cells might be applied to analyze mOGT caused cell apoptosis.

Key words： Human mOGT Matrix-targeting sequence Mitochondria fussion

Received: 02 November 2018 Published: 08 May 2019

ZTFLH:

Q819

Corresponding Authors: Hideki NAKANISHI E-mail: hideki@jiangnan.edu.cn

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Cite this article:

Feng LI,Xiao-dong GAO,Hideki NAKANISHI. Analysis of Martrix Targeting Sequence of Human Mitochondrial OGT in Saccharomyces cerevisiae. China Biotechnology, 2019, 39(4): 32-37.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20190405 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I4/32

Fig.1 The structure schematic of OGT

Table 1 Plasmids and primers

Fig.2 OGA could not rescue the growth defect of either mOGT or mOGT(H382A) expressing yeast cells

Fig.3 MOGT caused yeast growth defect depends on its MTS sequence

Fig.4 MOGT localized in yeast mitochondria (a)The mitochondria structure of yeast cell with vector (b)The mitochondria structure of yeast cell with mOGT-RFP (c)RFP signal of (b) (d)The overlayed image of (b) and (c) The mitochondria structure (green) and mOGT-RFP (red) localizations were represented by fluorescent microscope

Fig.5 MTS sequence caused yeast mitochondria fussion The mitochondria structure (green) and mOGT-RFP (red) localizations of yeast cell with vector (a), MTS15-RFP (b), MTS50-RFP (c) were represented by fluorescent microscope under bright field, red fluorescent, green fluorescent models (d)The overlayed image of red fluorescent and green fluorescent signals in (c)

Fig.6 MTS sequence caused yeast growth defect


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