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Article Figures & Data

Figures

  • Fig. 1.

    Restriction enzyme map of the AZU1-PRTN3-ELA2-ADN (APEA) locus. Cosmids used to identify the relative position of the ADN gene to the known genes in the locus are shown as thin horizontal lines above the restriction map. Restriction sites mapped are indicated. (B, BamHI; N, Not I; R, EcoRI; S, Sal I). Relative positions of the identified exons are shown for ADN (thick lines), and also for AZU1, PRTN3, and ELA2.

  • Fig. 2.

    Expression of the AZU1-PRTN3-ELA2-ADN cluster of genes in cell lines studied. Northern analysis was performed on U-937 cells, U-937 cells treated with 25 ng/mL TPA for 3 days (TPA), COLO 201 (COLO), HUT 78 (HUT), Jurkat, and K-562 cell lines and hybridized to32P-labeled human ELA2, AZU1, PRTN3, ADN, and human β-actin cDNA probes as indicated for each blot.

  • Fig. 3.

    Scale map of DHS in U-937 cells (vertical arrows) relative to the AZU1, PRTN3, ELA2, and ADN genes and to restriction fragments used for DHS mapping (short vertical lines), (A, Hpa I; B, BamHI; C, Sca I; G, Bgl II; L, Bcl I; M, Mlul; R, EcoRI; X, Xbal, and Y, BssHII). Horizontal arrows depict orientation of end-labeled detection of DHS. Exons from the 4 genes used as probes for DHS analysis are shown (half-boxes) for each restriction fragment used for DHS mapping. The EcoRI fragment encompassing AZU1 extends 24 kb beyond the AZU1 gene and the EcoRI fragment at the ADN end of the locus is 31 kb. Multiple analyses using different restriction fragments were used to confirm the location of the DHS in U-937 and in other cell lines.

  • Fig. 4.

    Analysis of chromatin structure at the AZU1-PRTN3-ELA2-ADN (APEA) locus in untreated U-937 cells. Nuclei were prepared and treated with increasing concentrations of DNase I (0 to 5 μg/mL, increasing from left to right in each case). The location of molecular weight DNA markers are shown on the left, in kilobases. DHS are indicated by solid arrowheads, with DHS-1 lying toward the AZU1 portion of the locus and DHS-15 at the ADN end of the locus. (A) DHS-1 to -4. DNase I–treated DNA was digested to completion with EcoRI and hybridized with a PCR amplified, 32P-labeled AZU1 exon 4/5 cDNA probe. (B) DHS-5 to -10. DNase I–treated DNA was digested to completion with Mlu I and hybridized with a PCR-amplified ELA2 exon I/II probe. (C) DHS-16 and -17. DNase I–treated DNA was digested to completion with Eco RI and hybridized with a 32P-labeled ADN cDNA probe. (D) DHS-11 and -12. DNase I–treated DNA was digested to completion with Bam HI and hybridized with a 32P-labeled ELA2 exon IV/V probe.

  • Fig. 5.

    Analysis of DHS-11 to -16 in COLO 201, U-937, and HL-60 cells. All lanes are DNase I–treated DNA digested to completion with Bgl II and hybridized with a 32P-labeled ELA2 exon IV/V cDNA probe. From left to right: C0, COLO 201 DNA not treated with DNase I; C1 to C3, COLO 201 DNA from nuclei treated with increasing concentrations of DNase I; H, HL-60 DNase I treated time-point; U, U-937 DNase I treated time-point. The locations of the DNA molecular weight markers used are shown on the left, and the location of DHS-11 to -16 are indicated by arrowheads on the right.

Tables

  • Table 1.

    Presence of DNase Hypersensitive Sites in Myeloid and Nonmyeloid Cell Lines

    1 2 3 4 5 6 7 8 9 10 1112 13 14 15 16 17
    U-937
    +TPA
    HL-60
    COLO
    HUT
    Jurkat
    K562
    PMN
    • The presence of DHS-1 to -17 (columns) are indicated by open arrows or thin arrows (extremely weak or barely distinguishable site) in each of the cell lines studied (rows). The presence of each DHS was confirmed by overlapping DHS analyses as for Fig 3.

    • Abbreviations: +TPA, TPA-treated U-937 cells; PMN, peripheral blood granulocytes.