Inhibition of Messenger RNA Transcriptional Activity in ML-1 Human Myeloblastic Leukemia Cell Nuclei by Antiserum to a c-myb-specific Peptide

Yoshio Honma, Motoo Hozumi, Jennifer D Black, Thomas Kieber-Emmons

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Antiserum to a synthetic peptide that defines a hydrophilic region within the putative c-myb translation product was prepared in the rabbit. In lysates from exponentially growing ML-1, human myeloblastic leukemia cells, the antiserum (“anti-myb”) reacted with five proteins of Mr58, 000, 75, 000, 85, 000, 90, 000 and 105, 000. Of these, only p75 and a trace of p85 were detected, by immunoblotting, in extracts derived from ML-1 cell nuclei. The proteins p58, p75 and p90 were present in readily detectable amounts only in the relatively immature myeloid cell lines ML-1 and HL-60, whereas in the more mature myeloid cell line THP-1 and in the lymphoid line BALL-1 only traces of these proteins were found. p85 and p105 were detected in lysates from all cell lines tested, including myeloid and lymphoid leukemia cells and mouse 3T3 cells. In lysates from ML-1 cells induced to differentiate to monocyte/macrophages or to granulocytes, the concentrations of p58 and p75 decreased in parallel with the cell population moving to maturity; in completely mature populations these two proteins were no longer detectable. In ML-1 cells arrested in G1by serum depletion, the amount of p58 and p75 and to a smaller extent that of p90 was decreased, whereas the concentration of p85 and p105 remained unchanged. In nuclei from exponentially growing ML-1 cells, the antiserum or its derived immunoglobulin fraction (“anti-myb IgG”) inhibited mRNA transcriptional activity by 30%. DNA synthesis was not affected. In contrast, in nuclei from differentiated ML-1 cells, the mRNA transcriptional activity was not significantly inhibited by anti-myb IgG. Similarly, in nuclei from ML-1 cells arrested largely in G1 by serum depletion for 2 days, mRNA transcriptional activity was inhibited by only 11%. Upon supplementation with serum, the mRNA transcriptional activity inhibitable by anti-myb IgG increased in parallel with the increasing rate of cell growth. The difference in total mRNA transcriptional activity observed in nuclei from cells of different growth rate was accounted for by the difference in transcriptional activity inhibitable by anti-myb IgG. This correlation between the rate of ML-1 cell growth, the concentration of a specific nuclear protein recognized by anti-myb IgG, and the extent of mRNA transcriptional activity inhibitable by the antibody suggests that a c-myb or c-myb-related product is involved in regulating the expression of some proliferation-related mRNAs.

Original languageEnglish (US)
Pages (from-to)1052-1057
Number of pages6
JournalCancer Research
Volume47
Issue number4
StatePublished - Jan 1 1987

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Cell Nucleus
Immune Sera
Leukemia
Messenger RNA
Peptides
Myeloid Cells
Cell Line
Proteins
Growth
Serum
Lymphoid Leukemia
3T3 Cells
Myeloid Leukemia
Nuclear Proteins
Immunoblotting
Granulocytes
Population
Immunoglobulins
Monocytes
Macrophages

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Inhibition of Messenger RNA Transcriptional Activity in ML-1 Human Myeloblastic Leukemia Cell Nuclei by Antiserum to a c-myb-specific Peptide. / Honma, Yoshio; Hozumi, Motoo; Black, Jennifer D; Kieber-Emmons, Thomas.

In: Cancer Research, Vol. 47, No. 4, 01.01.1987, p. 1052-1057.

Research output: Contribution to journalArticle

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abstract = "Antiserum to a synthetic peptide that defines a hydrophilic region within the putative c-myb translation product was prepared in the rabbit. In lysates from exponentially growing ML-1, human myeloblastic leukemia cells, the antiserum (“anti-myb”) reacted with five proteins of Mr58, 000, 75, 000, 85, 000, 90, 000 and 105, 000. Of these, only p75 and a trace of p85 were detected, by immunoblotting, in extracts derived from ML-1 cell nuclei. The proteins p58, p75 and p90 were present in readily detectable amounts only in the relatively immature myeloid cell lines ML-1 and HL-60, whereas in the more mature myeloid cell line THP-1 and in the lymphoid line BALL-1 only traces of these proteins were found. p85 and p105 were detected in lysates from all cell lines tested, including myeloid and lymphoid leukemia cells and mouse 3T3 cells. In lysates from ML-1 cells induced to differentiate to monocyte/macrophages or to granulocytes, the concentrations of p58 and p75 decreased in parallel with the cell population moving to maturity; in completely mature populations these two proteins were no longer detectable. In ML-1 cells arrested in G1by serum depletion, the amount of p58 and p75 and to a smaller extent that of p90 was decreased, whereas the concentration of p85 and p105 remained unchanged. In nuclei from exponentially growing ML-1 cells, the antiserum or its derived immunoglobulin fraction (“anti-myb IgG”) inhibited mRNA transcriptional activity by 30{\%}. DNA synthesis was not affected. In contrast, in nuclei from differentiated ML-1 cells, the mRNA transcriptional activity was not significantly inhibited by anti-myb IgG. Similarly, in nuclei from ML-1 cells arrested largely in G1 by serum depletion for 2 days, mRNA transcriptional activity was inhibited by only 11{\%}. Upon supplementation with serum, the mRNA transcriptional activity inhibitable by anti-myb IgG increased in parallel with the increasing rate of cell growth. The difference in total mRNA transcriptional activity observed in nuclei from cells of different growth rate was accounted for by the difference in transcriptional activity inhibitable by anti-myb IgG. This correlation between the rate of ML-1 cell growth, the concentration of a specific nuclear protein recognized by anti-myb IgG, and the extent of mRNA transcriptional activity inhibitable by the antibody suggests that a c-myb or c-myb-related product is involved in regulating the expression of some proliferation-related mRNAs.",
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N2 - Antiserum to a synthetic peptide that defines a hydrophilic region within the putative c-myb translation product was prepared in the rabbit. In lysates from exponentially growing ML-1, human myeloblastic leukemia cells, the antiserum (“anti-myb”) reacted with five proteins of Mr58, 000, 75, 000, 85, 000, 90, 000 and 105, 000. Of these, only p75 and a trace of p85 were detected, by immunoblotting, in extracts derived from ML-1 cell nuclei. The proteins p58, p75 and p90 were present in readily detectable amounts only in the relatively immature myeloid cell lines ML-1 and HL-60, whereas in the more mature myeloid cell line THP-1 and in the lymphoid line BALL-1 only traces of these proteins were found. p85 and p105 were detected in lysates from all cell lines tested, including myeloid and lymphoid leukemia cells and mouse 3T3 cells. In lysates from ML-1 cells induced to differentiate to monocyte/macrophages or to granulocytes, the concentrations of p58 and p75 decreased in parallel with the cell population moving to maturity; in completely mature populations these two proteins were no longer detectable. In ML-1 cells arrested in G1by serum depletion, the amount of p58 and p75 and to a smaller extent that of p90 was decreased, whereas the concentration of p85 and p105 remained unchanged. In nuclei from exponentially growing ML-1 cells, the antiserum or its derived immunoglobulin fraction (“anti-myb IgG”) inhibited mRNA transcriptional activity by 30%. DNA synthesis was not affected. In contrast, in nuclei from differentiated ML-1 cells, the mRNA transcriptional activity was not significantly inhibited by anti-myb IgG. Similarly, in nuclei from ML-1 cells arrested largely in G1 by serum depletion for 2 days, mRNA transcriptional activity was inhibited by only 11%. Upon supplementation with serum, the mRNA transcriptional activity inhibitable by anti-myb IgG increased in parallel with the increasing rate of cell growth. The difference in total mRNA transcriptional activity observed in nuclei from cells of different growth rate was accounted for by the difference in transcriptional activity inhibitable by anti-myb IgG. This correlation between the rate of ML-1 cell growth, the concentration of a specific nuclear protein recognized by anti-myb IgG, and the extent of mRNA transcriptional activity inhibitable by the antibody suggests that a c-myb or c-myb-related product is involved in regulating the expression of some proliferation-related mRNAs.

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