Cloning and characterization of cDNA encoding rat hemin-sensitive initiation factor-2α (eIF-2α) kinase. Evidence for multitissue expression

H. Mellor, K. M. Flowers, S. R. Kimball, L. S. Jefferson

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Abstract

Reticulocytes contain an eukaryotic initiation factor-2α (eIF-2α) kinase that is negatively regulated by hemin. This protein kinase, which has been termed heme-regulated inhibitor and heme-controlled repressor (HCR), has a strong inhibitory effect on the initiation of protein synthesis and plays an important role in translational control in these cells. Previous evidence has suggested that HCR may be an erythroid-specific enzyme. As reported herein, we have cloned and characterized the cDNA encoding an eIF-2α kinase from rat brain. The predicted amino acid sequence of the kinase from rat brain shows 82% homology to rabbit reticulocyte HCR with the greatest variation concentrated in a central region of approximately 135 amino acids located between protein kinase subdomains IV and VI. We have expressed the rat brain cDNA in Escherichia coli and have demonstrated that the recombinant enzyme is a hemin-sensitive eIF-2α kinase. We have also shown that mRNA recognized by the rat brain cDNA is expressed in a wide range of non-erythroid rat tissues at a lower but significant level compared with reticulocytes. These findings raise the possibility of additional, uncharacterized roles for HCR in non- erythroid cells.

Original languageEnglish (US)
Pages (from-to)10201-10204
Number of pages4
JournalJournal of Biological Chemistry
Volume269
Issue number14
StatePublished - Jan 1 1994

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Prokaryotic Initiation Factor-2
Eukaryotic Initiation Factor-2
Hemin
Cloning
eIF-2 Kinase
Rats
Organism Cloning
Phosphotransferases
Complementary DNA
Reticulocytes
Brain
Protein Kinases
Amino Acids
Erythroid Cells
Enzymes
Heme
Escherichia coli
Amino Acid Sequence
Tissue
Rabbits

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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abstract = "Reticulocytes contain an eukaryotic initiation factor-2α (eIF-2α) kinase that is negatively regulated by hemin. This protein kinase, which has been termed heme-regulated inhibitor and heme-controlled repressor (HCR), has a strong inhibitory effect on the initiation of protein synthesis and plays an important role in translational control in these cells. Previous evidence has suggested that HCR may be an erythroid-specific enzyme. As reported herein, we have cloned and characterized the cDNA encoding an eIF-2α kinase from rat brain. The predicted amino acid sequence of the kinase from rat brain shows 82{\%} homology to rabbit reticulocyte HCR with the greatest variation concentrated in a central region of approximately 135 amino acids located between protein kinase subdomains IV and VI. We have expressed the rat brain cDNA in Escherichia coli and have demonstrated that the recombinant enzyme is a hemin-sensitive eIF-2α kinase. We have also shown that mRNA recognized by the rat brain cDNA is expressed in a wide range of non-erythroid rat tissues at a lower but significant level compared with reticulocytes. These findings raise the possibility of additional, uncharacterized roles for HCR in non- erythroid cells.",
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AU - Mellor, H.

AU - Flowers, K. M.

AU - Kimball, S. R.

AU - Jefferson, L. S.

PY - 1994/1/1

Y1 - 1994/1/1

N2 - Reticulocytes contain an eukaryotic initiation factor-2α (eIF-2α) kinase that is negatively regulated by hemin. This protein kinase, which has been termed heme-regulated inhibitor and heme-controlled repressor (HCR), has a strong inhibitory effect on the initiation of protein synthesis and plays an important role in translational control in these cells. Previous evidence has suggested that HCR may be an erythroid-specific enzyme. As reported herein, we have cloned and characterized the cDNA encoding an eIF-2α kinase from rat brain. The predicted amino acid sequence of the kinase from rat brain shows 82% homology to rabbit reticulocyte HCR with the greatest variation concentrated in a central region of approximately 135 amino acids located between protein kinase subdomains IV and VI. We have expressed the rat brain cDNA in Escherichia coli and have demonstrated that the recombinant enzyme is a hemin-sensitive eIF-2α kinase. We have also shown that mRNA recognized by the rat brain cDNA is expressed in a wide range of non-erythroid rat tissues at a lower but significant level compared with reticulocytes. These findings raise the possibility of additional, uncharacterized roles for HCR in non- erythroid cells.

AB - Reticulocytes contain an eukaryotic initiation factor-2α (eIF-2α) kinase that is negatively regulated by hemin. This protein kinase, which has been termed heme-regulated inhibitor and heme-controlled repressor (HCR), has a strong inhibitory effect on the initiation of protein synthesis and plays an important role in translational control in these cells. Previous evidence has suggested that HCR may be an erythroid-specific enzyme. As reported herein, we have cloned and characterized the cDNA encoding an eIF-2α kinase from rat brain. The predicted amino acid sequence of the kinase from rat brain shows 82% homology to rabbit reticulocyte HCR with the greatest variation concentrated in a central region of approximately 135 amino acids located between protein kinase subdomains IV and VI. We have expressed the rat brain cDNA in Escherichia coli and have demonstrated that the recombinant enzyme is a hemin-sensitive eIF-2α kinase. We have also shown that mRNA recognized by the rat brain cDNA is expressed in a wide range of non-erythroid rat tissues at a lower but significant level compared with reticulocytes. These findings raise the possibility of additional, uncharacterized roles for HCR in non- erythroid cells.

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