AglH, a thermophilic UDP‑N‑acetylglucosamine‑1‑phosphate: dolichyl phosphate GlcNAc‑1‑phosphotransferase initiating protein N‑glycosylation pathway in Sulfolobus acidocaldarius, is capable of complementing the eukaryal Alg7

Benjamin H. Meyer (Lead / Corresponding author), Hosam Shams-Eldin, Sonja Verena Albers (Lead / Corresponding author)

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4 Citations (Scopus)
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Abstract

AglH, a predicted UDP-GlcNAc-1-phosphate: dolichyl phosphate GlcNAc-1-phosphotransferase, is initi-ating the protein N-glycosylation pathway in the thermoac-idophilic crenarchaeon Sulfolobus acidocaldarius. AglH successfully replaced the endogenous GlcNAc-1-phos-photransferase activity of Alg7 in a conditional lethal Sac-charomyces cerevisiae strain, in which the first step of the eukaryal protein N-glycosylation process was repressed. This study is one of the few examples of cross-domain complementation demonstrating a conserved polyprenyl phosphate transferase reaction within the eukaryal and archaeal domain like it was demonstrated for Methanococ-cus voltae (Shams-Eldin et al. 2008). The topology predic-tion and the alignment of the AglH membrane protein with GlcNAc-1-phosphotransferases from the three domains of life show significant conservation of amino acids within the different proposed cytoplasmic loops. Alanine mutations of selected conserved amino acids in the putative cytoplas-mic loops II (D100), IV (F220) and V (F264) demonstrated the importance of these amino acids for cross-domain AlgH activity in in vitro complementation assays in S. cerevisiae. Furthermore, antibiotic treatment interfering directly with the activity of dolichyl phosphate GlcNAc-1-phosphotrans-ferases confirmed the essentiality of N-glycosylation for cell survival.
Original languageEnglish
Pages (from-to)121-134
Number of pages14
JournalExtremophiles
Volume21
Issue number1
Early online date7 Nov 2016
DOIs
Publication statusPublished - Jan 2017
Event11th International Congress on Extremophiles - Kyoto, Japan
Duration: 12 Sep 201616 Sep 2016

Keywords

  • N-Glycosylation
  • Crenarchaea
  • Sulfolobus
  • Glycosylation
  • Dolichol phosphate
  • Alg7
  • Ag1H

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