Phosphorylation of mammalian translation initiation factor 5 (elF5) in vitro and in vivo

Romit Majumdar, Amitabha Bandyopadhyay, Haiteng Deng, Umadas Maitra

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Eukaryotic translation initiation factor 5 (elF5) interacts with the 40S initiation complex isolated a protein kinase from rabbit reticulocyte lysates on the basis of its ability to phosphorylate purified bacterially expressed recombinant rat elF5. Physical, biochemical and antigenic properties of this kinase identify it as casein kinase II (CK II). Mass spectrometric analysis of maximally in vitro phosphorylated elF5 localized the major phosphorylation sites at Ser-387 and Ser-388 near the C-terminus of elF5. These serine residues are embedded within a cluster of acidic amino acid residues and account for nearly 90% of the total in vitro elF5 phosphorylation. A minor phosphorylation site at Ser-174 was also observed. Alanine substitution mutagenesis at Ser-387 and Ser-388 of elF5 abolishes phosphorylation by the purified kinase as well as by crude reticulocyte lysates. The same mutations also abolish phosphorylation of elF5 when transfected into mammalian cells suggesting that CK II phosphorylates elF5 at these two serine residues in vivo as well.

Original languageEnglish (US)
Pages (from-to)1154-1162
Number of pages9
JournalNucleic Acids Research
Volume30
Issue number5
StatePublished - Mar 1 2002

Fingerprint

Peptide Initiation Factors
Phosphorylation
Casein Kinase II
Reticulocytes
Eukaryotic Initiation Factor-5
Serine
Phosphotransferases
Eukaryotic Initiation Factors
Acidic Amino Acids
Mutagenesis
Alanine
Protein Kinases
In Vitro Techniques
Rabbits
Mutation

ASJC Scopus subject areas

  • Genetics

Cite this

Majumdar, R., Bandyopadhyay, A., Deng, H., & Maitra, U. (2002). Phosphorylation of mammalian translation initiation factor 5 (elF5) in vitro and in vivo. Nucleic Acids Research, 30(5), 1154-1162.

Phosphorylation of mammalian translation initiation factor 5 (elF5) in vitro and in vivo. / Majumdar, Romit; Bandyopadhyay, Amitabha; Deng, Haiteng; Maitra, Umadas.

In: Nucleic Acids Research, Vol. 30, No. 5, 01.03.2002, p. 1154-1162.

Research output: Contribution to journalArticle

Majumdar, R, Bandyopadhyay, A, Deng, H & Maitra, U 2002, 'Phosphorylation of mammalian translation initiation factor 5 (elF5) in vitro and in vivo', Nucleic Acids Research, vol. 30, no. 5, pp. 1154-1162.
Majumdar R, Bandyopadhyay A, Deng H, Maitra U. Phosphorylation of mammalian translation initiation factor 5 (elF5) in vitro and in vivo. Nucleic Acids Research. 2002 Mar 1;30(5):1154-1162.
Majumdar, Romit ; Bandyopadhyay, Amitabha ; Deng, Haiteng ; Maitra, Umadas. / Phosphorylation of mammalian translation initiation factor 5 (elF5) in vitro and in vivo. In: Nucleic Acids Research. 2002 ; Vol. 30, No. 5. pp. 1154-1162.
@article{8a26e182166449e8911f62d21ae46329,
title = "Phosphorylation of mammalian translation initiation factor 5 (elF5) in vitro and in vivo",
abstract = "Eukaryotic translation initiation factor 5 (elF5) interacts with the 40S initiation complex isolated a protein kinase from rabbit reticulocyte lysates on the basis of its ability to phosphorylate purified bacterially expressed recombinant rat elF5. Physical, biochemical and antigenic properties of this kinase identify it as casein kinase II (CK II). Mass spectrometric analysis of maximally in vitro phosphorylated elF5 localized the major phosphorylation sites at Ser-387 and Ser-388 near the C-terminus of elF5. These serine residues are embedded within a cluster of acidic amino acid residues and account for nearly 90{\%} of the total in vitro elF5 phosphorylation. A minor phosphorylation site at Ser-174 was also observed. Alanine substitution mutagenesis at Ser-387 and Ser-388 of elF5 abolishes phosphorylation by the purified kinase as well as by crude reticulocyte lysates. The same mutations also abolish phosphorylation of elF5 when transfected into mammalian cells suggesting that CK II phosphorylates elF5 at these two serine residues in vivo as well.",
author = "Romit Majumdar and Amitabha Bandyopadhyay and Haiteng Deng and Umadas Maitra",
year = "2002",
month = "3",
day = "1",
language = "English (US)",
volume = "30",
pages = "1154--1162",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "5",

}

TY - JOUR

T1 - Phosphorylation of mammalian translation initiation factor 5 (elF5) in vitro and in vivo

AU - Majumdar, Romit

AU - Bandyopadhyay, Amitabha

AU - Deng, Haiteng

AU - Maitra, Umadas

PY - 2002/3/1

Y1 - 2002/3/1

N2 - Eukaryotic translation initiation factor 5 (elF5) interacts with the 40S initiation complex isolated a protein kinase from rabbit reticulocyte lysates on the basis of its ability to phosphorylate purified bacterially expressed recombinant rat elF5. Physical, biochemical and antigenic properties of this kinase identify it as casein kinase II (CK II). Mass spectrometric analysis of maximally in vitro phosphorylated elF5 localized the major phosphorylation sites at Ser-387 and Ser-388 near the C-terminus of elF5. These serine residues are embedded within a cluster of acidic amino acid residues and account for nearly 90% of the total in vitro elF5 phosphorylation. A minor phosphorylation site at Ser-174 was also observed. Alanine substitution mutagenesis at Ser-387 and Ser-388 of elF5 abolishes phosphorylation by the purified kinase as well as by crude reticulocyte lysates. The same mutations also abolish phosphorylation of elF5 when transfected into mammalian cells suggesting that CK II phosphorylates elF5 at these two serine residues in vivo as well.

AB - Eukaryotic translation initiation factor 5 (elF5) interacts with the 40S initiation complex isolated a protein kinase from rabbit reticulocyte lysates on the basis of its ability to phosphorylate purified bacterially expressed recombinant rat elF5. Physical, biochemical and antigenic properties of this kinase identify it as casein kinase II (CK II). Mass spectrometric analysis of maximally in vitro phosphorylated elF5 localized the major phosphorylation sites at Ser-387 and Ser-388 near the C-terminus of elF5. These serine residues are embedded within a cluster of acidic amino acid residues and account for nearly 90% of the total in vitro elF5 phosphorylation. A minor phosphorylation site at Ser-174 was also observed. Alanine substitution mutagenesis at Ser-387 and Ser-388 of elF5 abolishes phosphorylation by the purified kinase as well as by crude reticulocyte lysates. The same mutations also abolish phosphorylation of elF5 when transfected into mammalian cells suggesting that CK II phosphorylates elF5 at these two serine residues in vivo as well.

UR - http://www.scopus.com/inward/record.url?scp=0036493241&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036493241&partnerID=8YFLogxK

M3 - Article

C2 - 11861906

AN - SCOPUS:0036493241

VL - 30

SP - 1154

EP - 1162

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 5

ER -