Figure 2.
Figure 2. PML I isoform specifically interacts with AML1b. (A) Schematic representation of the PML protein and genomic structure of various PML transcripts. The PML protein contains the RBCC motif that consists of the RING finger domain (RING), the 2 B boxes (B1 and B2), and the helical coiled-coil region. The proline rich region (P) and serine-proline-rich region (S-P) have been thought to contain the putative phosphorylation sites.44 SUMOylation sites at lysines 65, 160, and 49056 are indicated. The PML gene consists of 9 exons (shaded boxes) with exon 7 divided into exons 7a and 7b, and the introns being retained in several transcripts as a consequence of alternative splicing (black boxes).54 The nomenclature for PML isoforms proposed by Jensen55 was used in this study. The alternative usage of 3′ exons generates multiple transcripts I-VI. In the case of PML III and PML VI, the encoding regions in which frame-shift occurred as a result of the retained intron are indicated by the gray color. The total number of amino acids for each isoform is given on the right side. (B) The interaction with AML1b is specific to the PML I isoform. BOSC23 cells were cotransfected with HA-tagged AML1b and together either with control vector (mock) or FLAG-tagged PML isoforms (I-VI). The expression of AML1 in the lysates of transfectants was detected by immunoblotting using anti-HA (3F10) antibody (top). The lysates of transfectants were immunoprecipitated with anti-FLAG (M2) antibody. The immunoprecipitates were analyzed by immunoblotting using anti-FLAG (M2) (middle) and anti-HA (3F10) antibodies (bottom). (C) Identification of regions of PML I required for interaction with AML1b. BOSC23 cells were cotransfected with FLAG-AML1b and either with control vector (mock) or constructs of PML I as indicated. The expression of PML I in the lysates of transfectants were detected by immunoblotting using anti-HA antibody (3F10) (top). The lysate of transfectants were immunoprecipitated with anti-FLAG (M2) antibody and were analyzed by immunoblotting using anti-FLAG (M2) (middle) and anti-HA (3F10) antibodies (bottom).

PML I isoform specifically interacts with AML1b. (A) Schematic representation of the PML protein and genomic structure of various PML transcripts. The PML protein contains the RBCC motif that consists of the RING finger domain (RING), the 2 B boxes (B1 and B2), and the helical coiled-coil region. The proline rich region (P) and serine-proline-rich region (S-P) have been thought to contain the putative phosphorylation sites.44  SUMOylation sites at lysines 65, 160, and 49056  are indicated. The PML gene consists of 9 exons (shaded boxes) with exon 7 divided into exons 7a and 7b, and the introns being retained in several transcripts as a consequence of alternative splicing (black boxes).54  The nomenclature for PML isoforms proposed by Jensen55  was used in this study. The alternative usage of 3′ exons generates multiple transcripts I-VI. In the case of PML III and PML VI, the encoding regions in which frame-shift occurred as a result of the retained intron are indicated by the gray color. The total number of amino acids for each isoform is given on the right side. (B) The interaction with AML1b is specific to the PML I isoform. BOSC23 cells were cotransfected with HA-tagged AML1b and together either with control vector (mock) or FLAG-tagged PML isoforms (I-VI). The expression of AML1 in the lysates of transfectants was detected by immunoblotting using anti-HA (3F10) antibody (top). The lysates of transfectants were immunoprecipitated with anti-FLAG (M2) antibody. The immunoprecipitates were analyzed by immunoblotting using anti-FLAG (M2) (middle) and anti-HA (3F10) antibodies (bottom). (C) Identification of regions of PML I required for interaction with AML1b. BOSC23 cells were cotransfected with FLAG-AML1b and either with control vector (mock) or constructs of PML I as indicated. The expression of PML I in the lysates of transfectants were detected by immunoblotting using anti-HA antibody (3F10) (top). The lysate of transfectants were immunoprecipitated with anti-FLAG (M2) antibody and were analyzed by immunoblotting using anti-FLAG (M2) (middle) and anti-HA (3F10) antibodies (bottom).

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