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A glutamine/asparagine-rich fragment of Gln3, but not the full-length protein, aggregates in Saccharomyces cerevisiae

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Abstract

The amino acid sequence of protein Gln3 in yeast Saccharomyces cerevisiae has a region enriched with Gln (Q) and Asn (N) residues. In this study, we analyzed the effects of overexpression of Gln3 and its Q/N-rich fragment fused with yellow fluorescent protein (YFP). Being overexpressed, full-length Gln3-YFP does not form aggregates, inhibits vegetative growth, and demonstrates nuclear localization, while the Q/N-rich fragment (Gln3QN) fused with YFP forms aggregates that do not colocalize with the nucleus and do not affect growth of the cells. Although detergent-resistant aggregates of Gln3QN are formed in the absence of yeast prions, the aggregation of Gln3QN significantly increases in the presence of [PIN +] prion, while in the presence of two prions, [PSI +] and [PIN +], the percentage of cells with Gln3QN aggregates is significantly lower than in the strain bearing only [PIN +]. Data on colocalization demonstrate that this effect is mediated by interaction between Gln3QN aggregates and [PSI +] and [PIN +] prions.

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Abbreviations

CFP:

cyan fluorescent protein

DAPI:

4′,6-diamidino-2-phenylindole (a fluorescent dye specific to AT-rich regions of DNA)

Gln3QN:

asparagine-glutamine-rich fragment of Gln3 protein (a.a. 166-242)

[PIN +]:

prion isoform of Rnq1 protein

[PSI +]:

prion isoform of Sup35 protein

SDDAGE:

Semi-Denaturing Detergent Agarose Gel Electrophoresis

Sup35NM:

prion-forming region of Sup35 protein lacking the C-terminal domain functioning as a translation release factor

YFP:

yellow fluorescent protein

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Authors and Affiliations

  1. Department of Genetics and Biotechnology, St. Petersburg State University, 199034, St. Petersburg, Russia

    K. S. Antonets, H. M. Sargsyan & A. A. Nizhnikov

  2. Vavilov Institute of General Genetics, St. Petersburg Branch, Russian Academy of Sciences, 199034, St. Petersburg, Russia

    K. S. Antonets & A. A. Nizhnikov

  3. All-Russia Research Institute for Agricultural Microbiology, 196608 Pushkin, St. Petersburg, Russia

    A. A. Nizhnikov

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  1. K. S. Antonets
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  2. H. M. Sargsyan
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Correspondence to A. A. Nizhnikov.

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Original Russian Text © K. S. Antonets, H. M. Sargsyan, A. A. Nizhnikov, 2016, published in Biokhimiya, 2016, Vol. 81, No. 4, pp. 555-562.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM15-341, January 31, 2016.

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Antonets, K.S., Sargsyan, H.M. & Nizhnikov, A.A. A glutamine/asparagine-rich fragment of Gln3, but not the full-length protein, aggregates in Saccharomyces cerevisiae . Biochemistry Moscow 81, 407–413 (2016). https://doi.org/10.1134/S0006297916040118

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