Ribosomal protein S3 (rpS3) secreted from various cancer cells is N-linked glycosylated

Yong Joong Kim, Min Seon Lee, Hag Dong Kim, Joon Kim

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Ribosomal protein S3 (rpS3) is a 243 amino acid component of the 40S ribosomal small subunit. It has multiple roles in translation and extra-ribosomal functions like apoptosis and DNA repair. RpS3 is secreted only in cancer cell lines. Presently, mass spectrometry analysis revealed rpS3 to be glycosylated at the Asn165 residue. A point mutation at this residue decreased secretion of rpS3 in cancer cell lines. Secretion was also inhibited by the endoplasmic reticulum (ER)-Golgi transport inhibitor Brefeldin A and by Tunicamycin, an inhibitor of N-linked glycosylation. N-linked glycosylation of rpS3 was confirmed as necessary for rpS3 secretion into culture media via the ER-Golgi dependent pathway. RpS3 bound to Concanavalin A, a carbohydrate binding lectin protein, while treatment with peptide-N-glycosidase F shifted the secreted rpS3 to a lower molecular weight band. In addition, the N165G mutant of rpS3 displayed reduced secretion compared to the wild-type. An in vitro binding assay detected rpS3 homodimer formation via the N-terminal region (rpS3:1-85) and a middle region (rpS3:95-158). The results indicate that the Asn 165 residue of rpS3 is a critical site for N-linked glycosylation and passage through the ER-Golgi secretion pathway.

Original languageEnglish
Pages (from-to)80350-80362
Number of pages13
JournalOncotarget
Volume7
Issue number49
DOIs
Publication statusPublished - 2016

Fingerprint

Neoplasms
Glycosylation
Endoplasmic Reticulum
ribosomal protein S3
Eukaryotic Small Ribosome Subunits
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Brefeldin A
Tunicamycin
Cell Line
Secretory Pathway
Mutant Proteins
Concanavalin A
Point Mutation
Lectins
DNA Repair
Culture Media
Mass Spectrometry
Molecular Weight
Apoptosis
Amino Acids

Keywords

  • Glycosylation
  • Ribosomal protein S3
  • Ribosome
  • Secretion

ASJC Scopus subject areas

  • Oncology

Cite this

Ribosomal protein S3 (rpS3) secreted from various cancer cells is N-linked glycosylated. / Kim, Yong Joong; Lee, Min Seon; Kim, Hag Dong; Kim, Joon.

In: Oncotarget, Vol. 7, No. 49, 2016, p. 80350-80362.

Research output: Contribution to journalArticle

Kim, Yong Joong ; Lee, Min Seon ; Kim, Hag Dong ; Kim, Joon. / Ribosomal protein S3 (rpS3) secreted from various cancer cells is N-linked glycosylated. In: Oncotarget. 2016 ; Vol. 7, No. 49. pp. 80350-80362.
@article{d044d19f9613422e9a1eaed85ca60e58,
title = "Ribosomal protein S3 (rpS3) secreted from various cancer cells is N-linked glycosylated",
abstract = "Ribosomal protein S3 (rpS3) is a 243 amino acid component of the 40S ribosomal small subunit. It has multiple roles in translation and extra-ribosomal functions like apoptosis and DNA repair. RpS3 is secreted only in cancer cell lines. Presently, mass spectrometry analysis revealed rpS3 to be glycosylated at the Asn165 residue. A point mutation at this residue decreased secretion of rpS3 in cancer cell lines. Secretion was also inhibited by the endoplasmic reticulum (ER)-Golgi transport inhibitor Brefeldin A and by Tunicamycin, an inhibitor of N-linked glycosylation. N-linked glycosylation of rpS3 was confirmed as necessary for rpS3 secretion into culture media via the ER-Golgi dependent pathway. RpS3 bound to Concanavalin A, a carbohydrate binding lectin protein, while treatment with peptide-N-glycosidase F shifted the secreted rpS3 to a lower molecular weight band. In addition, the N165G mutant of rpS3 displayed reduced secretion compared to the wild-type. An in vitro binding assay detected rpS3 homodimer formation via the N-terminal region (rpS3:1-85) and a middle region (rpS3:95-158). The results indicate that the Asn 165 residue of rpS3 is a critical site for N-linked glycosylation and passage through the ER-Golgi secretion pathway.",
keywords = "Glycosylation, Ribosomal protein S3, Ribosome, Secretion",
author = "Kim, {Yong Joong} and Lee, {Min Seon} and Kim, {Hag Dong} and Joon Kim",
year = "2016",
doi = "10.18632/oncotarget.10180",
language = "English",
volume = "7",
pages = "80350--80362",
journal = "Oncotarget",
issn = "1949-2553",
publisher = "Impact Journals",
number = "49",

}

TY - JOUR

T1 - Ribosomal protein S3 (rpS3) secreted from various cancer cells is N-linked glycosylated

AU - Kim, Yong Joong

AU - Lee, Min Seon

AU - Kim, Hag Dong

AU - Kim, Joon

PY - 2016

Y1 - 2016

N2 - Ribosomal protein S3 (rpS3) is a 243 amino acid component of the 40S ribosomal small subunit. It has multiple roles in translation and extra-ribosomal functions like apoptosis and DNA repair. RpS3 is secreted only in cancer cell lines. Presently, mass spectrometry analysis revealed rpS3 to be glycosylated at the Asn165 residue. A point mutation at this residue decreased secretion of rpS3 in cancer cell lines. Secretion was also inhibited by the endoplasmic reticulum (ER)-Golgi transport inhibitor Brefeldin A and by Tunicamycin, an inhibitor of N-linked glycosylation. N-linked glycosylation of rpS3 was confirmed as necessary for rpS3 secretion into culture media via the ER-Golgi dependent pathway. RpS3 bound to Concanavalin A, a carbohydrate binding lectin protein, while treatment with peptide-N-glycosidase F shifted the secreted rpS3 to a lower molecular weight band. In addition, the N165G mutant of rpS3 displayed reduced secretion compared to the wild-type. An in vitro binding assay detected rpS3 homodimer formation via the N-terminal region (rpS3:1-85) and a middle region (rpS3:95-158). The results indicate that the Asn 165 residue of rpS3 is a critical site for N-linked glycosylation and passage through the ER-Golgi secretion pathway.

AB - Ribosomal protein S3 (rpS3) is a 243 amino acid component of the 40S ribosomal small subunit. It has multiple roles in translation and extra-ribosomal functions like apoptosis and DNA repair. RpS3 is secreted only in cancer cell lines. Presently, mass spectrometry analysis revealed rpS3 to be glycosylated at the Asn165 residue. A point mutation at this residue decreased secretion of rpS3 in cancer cell lines. Secretion was also inhibited by the endoplasmic reticulum (ER)-Golgi transport inhibitor Brefeldin A and by Tunicamycin, an inhibitor of N-linked glycosylation. N-linked glycosylation of rpS3 was confirmed as necessary for rpS3 secretion into culture media via the ER-Golgi dependent pathway. RpS3 bound to Concanavalin A, a carbohydrate binding lectin protein, while treatment with peptide-N-glycosidase F shifted the secreted rpS3 to a lower molecular weight band. In addition, the N165G mutant of rpS3 displayed reduced secretion compared to the wild-type. An in vitro binding assay detected rpS3 homodimer formation via the N-terminal region (rpS3:1-85) and a middle region (rpS3:95-158). The results indicate that the Asn 165 residue of rpS3 is a critical site for N-linked glycosylation and passage through the ER-Golgi secretion pathway.

KW - Glycosylation

KW - Ribosomal protein S3

KW - Ribosome

KW - Secretion

UR - http://www.scopus.com/inward/record.url?scp=85001094040&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85001094040&partnerID=8YFLogxK

U2 - 10.18632/oncotarget.10180

DO - 10.18632/oncotarget.10180

M3 - Article

C2 - 27384988

AN - SCOPUS:85001094040

VL - 7

SP - 80350

EP - 80362

JO - Oncotarget

JF - Oncotarget

SN - 1949-2553

IS - 49

ER -