Myelomonocytic cell lineage expression of the neutrophil elastase gene

H. Takahashi, T. Nukiwa, P. Basset, Ronald Crystal

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

Human neutrophil elastase (NE) functions as a powerful serine protease capable of attacking a broad range of proteins. To examine the cellular site(s) of NE gene expression, a 0.65-kilobase cDNA (pPB15) complementary to the coding region of the NE gene was cloned from the cell line U937 using an oligonucleotide based on the known NE protein sequence. The sequence of pPB15 demonstrated that it coded for the 173 C-terminal residues of the 218 amino acids that comprise the mature NE protein, plus an additional 3' 60 base pairs prior to the in-frame stop codon, suggesting the NE mRNA contains sequences for a 20-residue C-terminal 'pro' peptide that is not found in the mature protein. Northern analysis using 32P-labeled pPB15 as a probe revealed that neutrophils do not contain detectable NE mRNA transcripts despite the fact that this cell carries large amounts of this protein. Furthermore, resting and activated blood monocytes also contained no detectable NE mRNA transcripts, although these cells also carry detectable NE. In contrast, bone marrow precursor cells contained NE transcripts, suggesting the NE gene is expressed in blood precursor cells. In this regard, evaluation of HL-60 cells, a human cell line with myelomonocytic lineage features, demonstrated NE transcripts in resting cells and increased NE mRNA levels when the cells were induced toward the myelocytic lineage with dimethyl sulfoxide. However, when the HL-60 cells were induced toward the monocytic lineage with phorbol 12-myristate 13-acetate, NE transcripts were lost even though transcripts for interleukin-1β were plentiful. Together, these observations are consistent with the concept that the NE gene is not expressed in the blood cells that carry the protein, but in bone marrow precursors that express NE transcripts about the time of commitment to the myelocytic series.

Original languageEnglish
Pages (from-to)2543-2547
Number of pages5
JournalJournal of Biological Chemistry
Volume263
Issue number5
Publication statusPublished - 1 Jan 1988
Externally publishedYes

Fingerprint

Leukocyte Elastase
Cell Lineage
Genes
Messenger RNA
Proteins
Blood
HL-60 Cells
Cells
Blood Cells
Bone
Cell Line
Terminator Codon
Serine Proteases
Dimethyl Sulfoxide
Interleukin-1

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Myelomonocytic cell lineage expression of the neutrophil elastase gene. / Takahashi, H.; Nukiwa, T.; Basset, P.; Crystal, Ronald.

In: Journal of Biological Chemistry, Vol. 263, No. 5, 01.01.1988, p. 2543-2547.

Research output: Contribution to journalArticle

Takahashi, H, Nukiwa, T, Basset, P & Crystal, R 1988, 'Myelomonocytic cell lineage expression of the neutrophil elastase gene', Journal of Biological Chemistry, vol. 263, no. 5, pp. 2543-2547.
Takahashi, H. ; Nukiwa, T. ; Basset, P. ; Crystal, Ronald. / Myelomonocytic cell lineage expression of the neutrophil elastase gene. In: Journal of Biological Chemistry. 1988 ; Vol. 263, No. 5. pp. 2543-2547.
@article{5bc9e833f6fa4dd6932bd39bd7a1286b,
title = "Myelomonocytic cell lineage expression of the neutrophil elastase gene",
abstract = "Human neutrophil elastase (NE) functions as a powerful serine protease capable of attacking a broad range of proteins. To examine the cellular site(s) of NE gene expression, a 0.65-kilobase cDNA (pPB15) complementary to the coding region of the NE gene was cloned from the cell line U937 using an oligonucleotide based on the known NE protein sequence. The sequence of pPB15 demonstrated that it coded for the 173 C-terminal residues of the 218 amino acids that comprise the mature NE protein, plus an additional 3' 60 base pairs prior to the in-frame stop codon, suggesting the NE mRNA contains sequences for a 20-residue C-terminal 'pro' peptide that is not found in the mature protein. Northern analysis using 32P-labeled pPB15 as a probe revealed that neutrophils do not contain detectable NE mRNA transcripts despite the fact that this cell carries large amounts of this protein. Furthermore, resting and activated blood monocytes also contained no detectable NE mRNA transcripts, although these cells also carry detectable NE. In contrast, bone marrow precursor cells contained NE transcripts, suggesting the NE gene is expressed in blood precursor cells. In this regard, evaluation of HL-60 cells, a human cell line with myelomonocytic lineage features, demonstrated NE transcripts in resting cells and increased NE mRNA levels when the cells were induced toward the myelocytic lineage with dimethyl sulfoxide. However, when the HL-60 cells were induced toward the monocytic lineage with phorbol 12-myristate 13-acetate, NE transcripts were lost even though transcripts for interleukin-1β were plentiful. Together, these observations are consistent with the concept that the NE gene is not expressed in the blood cells that carry the protein, but in bone marrow precursors that express NE transcripts about the time of commitment to the myelocytic series.",
author = "H. Takahashi and T. Nukiwa and P. Basset and Ronald Crystal",
year = "1988",
month = "1",
day = "1",
language = "English",
volume = "263",
pages = "2543--2547",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "5",

}

TY - JOUR

T1 - Myelomonocytic cell lineage expression of the neutrophil elastase gene

AU - Takahashi, H.

AU - Nukiwa, T.

AU - Basset, P.

AU - Crystal, Ronald

PY - 1988/1/1

Y1 - 1988/1/1

N2 - Human neutrophil elastase (NE) functions as a powerful serine protease capable of attacking a broad range of proteins. To examine the cellular site(s) of NE gene expression, a 0.65-kilobase cDNA (pPB15) complementary to the coding region of the NE gene was cloned from the cell line U937 using an oligonucleotide based on the known NE protein sequence. The sequence of pPB15 demonstrated that it coded for the 173 C-terminal residues of the 218 amino acids that comprise the mature NE protein, plus an additional 3' 60 base pairs prior to the in-frame stop codon, suggesting the NE mRNA contains sequences for a 20-residue C-terminal 'pro' peptide that is not found in the mature protein. Northern analysis using 32P-labeled pPB15 as a probe revealed that neutrophils do not contain detectable NE mRNA transcripts despite the fact that this cell carries large amounts of this protein. Furthermore, resting and activated blood monocytes also contained no detectable NE mRNA transcripts, although these cells also carry detectable NE. In contrast, bone marrow precursor cells contained NE transcripts, suggesting the NE gene is expressed in blood precursor cells. In this regard, evaluation of HL-60 cells, a human cell line with myelomonocytic lineage features, demonstrated NE transcripts in resting cells and increased NE mRNA levels when the cells were induced toward the myelocytic lineage with dimethyl sulfoxide. However, when the HL-60 cells were induced toward the monocytic lineage with phorbol 12-myristate 13-acetate, NE transcripts were lost even though transcripts for interleukin-1β were plentiful. Together, these observations are consistent with the concept that the NE gene is not expressed in the blood cells that carry the protein, but in bone marrow precursors that express NE transcripts about the time of commitment to the myelocytic series.

AB - Human neutrophil elastase (NE) functions as a powerful serine protease capable of attacking a broad range of proteins. To examine the cellular site(s) of NE gene expression, a 0.65-kilobase cDNA (pPB15) complementary to the coding region of the NE gene was cloned from the cell line U937 using an oligonucleotide based on the known NE protein sequence. The sequence of pPB15 demonstrated that it coded for the 173 C-terminal residues of the 218 amino acids that comprise the mature NE protein, plus an additional 3' 60 base pairs prior to the in-frame stop codon, suggesting the NE mRNA contains sequences for a 20-residue C-terminal 'pro' peptide that is not found in the mature protein. Northern analysis using 32P-labeled pPB15 as a probe revealed that neutrophils do not contain detectable NE mRNA transcripts despite the fact that this cell carries large amounts of this protein. Furthermore, resting and activated blood monocytes also contained no detectable NE mRNA transcripts, although these cells also carry detectable NE. In contrast, bone marrow precursor cells contained NE transcripts, suggesting the NE gene is expressed in blood precursor cells. In this regard, evaluation of HL-60 cells, a human cell line with myelomonocytic lineage features, demonstrated NE transcripts in resting cells and increased NE mRNA levels when the cells were induced toward the myelocytic lineage with dimethyl sulfoxide. However, when the HL-60 cells were induced toward the monocytic lineage with phorbol 12-myristate 13-acetate, NE transcripts were lost even though transcripts for interleukin-1β were plentiful. Together, these observations are consistent with the concept that the NE gene is not expressed in the blood cells that carry the protein, but in bone marrow precursors that express NE transcripts about the time of commitment to the myelocytic series.

UR - http://www.scopus.com/inward/record.url?scp=0023856438&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023856438&partnerID=8YFLogxK

M3 - Article

VL - 263

SP - 2543

EP - 2547

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 5

ER -