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)

Research output: Contribution to journalSpecial issue

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

Fingerprint

Sulfolobus acidocaldarius
Glycosylation
Amino Acids
Proteins
Phosphotransferases
Uridine Diphosphate
Transferases
Alanine
Saccharomyces cerevisiae
Cell Survival
Membrane Proteins
Phosphates
Anti-Bacterial Agents
Mutation
N-acetylglucopyranosylamine
dolichol monophosphate

Keywords

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

Cite this

@article{d703de4264524be285dde111d86643c4,
title = "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",
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.",
keywords = "N-Glycosylation, Crenarchaea, Sulfolobus, Glycosylation, Dolichol phosphate, Alg7, Ag1H",
author = "Meyer, {Benjamin H.} and Hosam Shams-Eldin and Albers, {Sonja Verena}",
note = "BHM was supported by an ERC starting grant (311523, ARCHAELLUM). This work was supported by intramural funds of the Max Planck Society to SVA.",
year = "2017",
month = "1",
doi = "10.1007/s00792-016-0890-2",
language = "English",
volume = "21",
pages = "121--134",
journal = "Extremophiles",
issn = "1431-0651",
publisher = "Springer Verlag",
number = "1",

}

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

In: Extremophiles, Vol. 21, No. 1, 01.2017, p. 121-134.

Research output: Contribution to journalSpecial issue

TY - JOUR

T1 - AglH, a thermophilic UDP‑N‑acetylglucosamine‑1‑phosphate

T2 - dolichyl phosphate GlcNAc‑1‑phosphotransferase initiating protein N‑glycosylation pathway in Sulfolobus acidocaldarius, is capable of complementing the eukaryal Alg7

AU - Meyer, Benjamin H.

AU - Shams-Eldin, Hosam

AU - Albers, Sonja Verena

N1 - BHM was supported by an ERC starting grant (311523, ARCHAELLUM). This work was supported by intramural funds of the Max Planck Society to SVA.

PY - 2017/1

Y1 - 2017/1

N2 - 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.

AB - 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.

KW - N-Glycosylation

KW - Crenarchaea

KW - Sulfolobus

KW - Glycosylation

KW - Dolichol phosphate

KW - Alg7

KW - Ag1H

U2 - 10.1007/s00792-016-0890-2

DO - 10.1007/s00792-016-0890-2

M3 - Special issue

C2 - 27822701

VL - 21

SP - 121

EP - 134

JO - Extremophiles

JF - Extremophiles

SN - 1431-0651

IS - 1

ER -