The analysis of the polyamine oxidase genes in the methylotrophic yeast Komagataella phaffii

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Abstract

Polyamines are present in all living cells and regulate a wide range of biological processes. In Saccharomyces cerevisiae the polyamine oxidase Fms1p converts spermine to spermidine and 3-aminopropionaldehyde, which is necessary for the synthesis of pantothenic acid and hypusination. This paper shows that S. cerevisiae FMS1 gene orthologs are present in all major representatives of the Saccharomycotina subdivision, but their copy numbers are different. In the Komagataella phaffii (Pichia pastoris) yeast, two polyamine oxidase genes (KpFMS1 and KpFMS2) were identified, and the regulation of their promoters activity was studied.

About the authors

Alina V. Ivanova

St. Petersburg State University

Email: alinalans@gmail.com

4th year Student of the Department of Genetics and Biotechnology

Russian Federation, 7/9, Universitetskaya embankment, Saint-Petersburg, 199034

Anton V. Sidorin

St. Petersburg State University

Email: spacerocketpilot@gmail.com

Bachelor of Science (BSc) in the Genetics and Biotechnology Department

Russian Federation, 7/9, Universitetskaya embankment, Saint-Petersburg, 199034

Elena V. Sambuk

St. Petersburg State University

Email: esambuk@mail.ru
ORCID iD: 0000-0003-0837-0498
SPIN-code: 8281-8020
Scopus Author ID: 6603061322
ResearcherId: H-6895-2013

Doctor of Science, Professor of the Department of Genetics and Biotechnology

Russian Federation, 7/9, Universitetskaya embankment, Saint-Petersburg, 199034

Andrei M. Rumyantsev

St. Petersburg State University

Author for correspondence.
Email: rumyantsev-am@mail.ru
ORCID iD: 0000-0002-1744-3890
SPIN-code: 9335-1184
Scopus Author ID: 55370658800

PhD, Senior Researcher of the Department of Genetics and Biotechnology

Russian Federation, 7/9, Universitetskaya embankment, Saint-Petersburg, 199034

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Supplementary files

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2. Table 1. The primer sequences used in the work

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3. Fig. 1. The prevalence of polyamine oxidases among the main representatives of the Saccharomycotina subdivision. Phylogenetic relationships are shown based on the results obtained in [21]. WGD refers to the whole-genomic duplication that the ancestors of S. cerevisiae and related species underwent [28]

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4. Fig. 2. The results of multiple comparisons of polyamine oxidases amino acid sequences from the studied yeast species. The table shows the PTS1 sequences at the C-terminus of polyamine oxidases, which provide the peroxisomal localization of proteins. The underlined amino acids differ from the consensus sequence known for S. cerevisiae [22]

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5. Fig. 3. The arrangement of KpFMS2 and AOX1 K. phaffii genes. The binding sites of the transcription factors Mxr1p and Nrg1p (the main regulators of AOX1 gene) are shown

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6. Fig. 4. The activity of reporter acid phosphatase synthesized by K. phaffii PFMS1-4-GS115, PFMS2-4-GS115 and tr2-4-GS115 strains during their growth on the media with different carbon and nitrogen sources

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Copyright (c) 2019 Ivanova A.V., Sidorin A.V., Sambuk E.V., Rumyantsev A.M.

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This work is licensed under a Creative Commons Attribution 4.0 International License.
 


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