Plzf regulates limb and axial skeletal patterning

Abstract

The promyelocytic leukaemia zinc finger (Plzf) protein (encoded by the gene Zfp145) belongs to the POZ/zinc-finger family of transcription factors. Here we generate Zfp145−/− mice and show that Plzf is essential for patterning of the limb and axial skeleton. Plzf inactivation results in patterning defects affecting all skeletal structures of the limb, including homeotic transformations of anterior skeletal elements into posterior structures. We demonstrate that Plzf acts as a growth-inhibitory and pro-apoptotic factor in the limb bud. The expression of members of the abdominal b (Abdb) Hox gene complex, as well as genes encoding bone morphogenetic proteins (Bmps), is altered in the developing limb of Zfp145−/− mice. Plzf regulates the expression of these genes in the absence of aberrant polarizing activity and independently of known patterning genes. Zfp145−/− mice also exhibit anterior-directed homeotic transformation throughout the axial skeleton with associated alterations in Hox gene expression. Plzf is therefore a mediator of anterior-to-posterior (AP) patterning in both the axial and appendicular skeleton and acts as a regulator of Hox gene expression.

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Figure 1: Targeted disruption of Zfp145.
Figure 2: Autopod skeleton.
Figure 3: Zeugopod skeleton.
Figure 4: Increased proliferation and decreased apoptosis in the hindlimb of Zfp145−/− mice.
Figure 5: Expression of Zfp145 in the hindlimb.
Figure 6: Altered Hox and Bmp gene expression in the hindlimb of Zfp145−/− mice.
Figure 7: Homeotic transformations and patterning defects in the axial skeleton of Zfp145−/− mice.
Figure 8: Expression of Hox genes within prevertebrae (Pv).

References

  1. 1

    Pearse, R.V. & Tabin, C.J. The molecular ZPA. J. Exp. Zoology 282, 677–690 (1998).

  2. 2

    Krumlauf, R. Hox genes in vertebrate development. Cell 78, 191–201 (1994).

  3. 3

    Chen, Z. et al. Fusion between a novel Krüppel-like zinc finger gene and the retinoic acid receptor-α locus due to a variant t(11;17) translocation associated with acute promyelocytic leukaemia. EMBO J. 12, 1161–1167 (1995).

  4. 4

    Li, J.Y. et al. Sequence-specific DNA binding and transcriptional regulation by the promyelocytic leukemia zinc finger protein. J. Biol. Chem. 272, 22447–22455 (1997).

  5. 5

    Reid, A. et al. Leukemia translocation gene, PLZF, is expressed with a speckled nuclear pattern in early hematopoietic progenitors. Blood 86, 4544–4552 (1995).

  6. 6

    Chang, C.C., Ye, B.H., Chaganti, R.S. & Dalla-Favera, R. BCL-6, a POZ/zinc-finger protein, is a sequence-specific transcriptional repressor. Proc. Natl Acad. Sci. USA 93, 6947–6952 (1996).

  7. 7

    Hong, S.H., David, G., Wong, C.W., Dejean, A. & Privalsky, M.L. SMRT corepressor interacts with PLZF and with the PML-retinoic acid receptor-α (RARα) and PLZF-RARα oncoproteins associated with acute promyelocytic leukemia. Proc. Natl Acad. Sci. USA 94, 9028–9033 (1997).

  8. 8

    He, L.-Z. et al. Distinct interactions of PML-RARα and PLZF-RARα with transcriptional co-repressors determine differential responses to retinoic acid in APL. Nature Genet. 18, 126–135 (1998).

  9. 9

    Tickle, C. The number of polarizing region cells required to specify additional digits in the developing chick wing. Nature 289, 295–298 (1981).

  10. 10

    Riddle, R.D., Johnson, E.L. & Tabin, C. Sonic Hedgehog mediates the polarizing activity of the ZPA. Cell 75, 1401–1416 (1993).

  11. 11

    Morgan, B.A., Izpisua-Belmonte, J.C., Duboule, D. & Tabin, C.J. Targeted misexpression of Hox-4.6 in the avian limb bud causes apparent homeotic transformations. Nature 358, 236–239 (1992).

  12. 12

    Goff, D.J. & Tabin, C.J. Analysis of Hoxd-13 and Hoxd-11 misexpression in chick limb buds reveals that Hox genes affect both bone condensation and growth. Development 124, 627–636 (1997).

  13. 13

    Knezevic, V. et al. Hoxd-12 differentially affects preaxial and postaxial chondrogenic branches in the limb and regulates Sonic hedgehog in a positive feedback loop. Development 124, 4523–4536 (1997).

  14. 14

    Small, K.M. & Potter, S.S. Homeotic transformations and limb defects in HoxA11 mutant mice. Genes Dev. 7, 2318–2328 (1993).

  15. 15

    Yokouchi, Y. et al. Misexpression of Hoxa-13 induces cartilage homeotic transformation and changes cell adhesiveness in chick limb buds. Genes Dev. 9, 2509–2522 (1995).

  16. 16

    Fromental-Ramin, C. et al. Hoxa-13 and Hoxd-13 play a crucial role in the patterning of the limb autopod. Development 122, 2997–3011 (1996).

  17. 17

    Davis, A.P., Witte, D.P., Hsieh-Li, H.M., Potter, S.S. & Capecchi, M.R. Absence of radius and ulna in mice lacking hoxa-11 and hoxd-11. Nature 375, 791–795 (1995).

  18. 18

    Zou, H. & Niswander, L. Requirement for BMP signaling in interdigital apoptosis and scale formation. Science 272, 738–741 (1996).

  19. 19

    Pollack, R.A., Jay, G. & Bieberich, C.J. Altering the boundaries of Hox3.1 expression: evidence for antipodal gene expression. Cell 71, 911–923 (1992).

  20. 20

    Jegalian, B.G. & Robertis, E.M. Homeotic transformations in the mouse induced by overexpression of a human Hox3.3 transgene. Cell 71, 901–910 (1992).

  21. 21

    Avantaggiato, V. et al. Developmental analysis of murine promyelocyte leukemia zinc finger (PLZF) gene expression: implications for the neuromeric model of the forebrain organization. J. Neurosci. 15, 4927–4942 (1995).

  22. 22

    Luo, G. et al. BMP-7 is an inducer of nephrogenesis, and is also required for eye development and skeletal patterning. Genes Dev. 9, 2808–2820 (1995).

  23. 23

    Dudley, A.T., Lyons, K.M. & Robertson, E.J.A. Requirement for bone morphogenetic protein-7 during development of the mammalian kidney and eye. Genes Dev. 9, 2795–2807 (1995).

  24. 24

    Hoatlin, M.E. et al. A novel BTB/POZ transcriptional repressor protein interacts with the Fanconi anemia group C protein and PLZF. Blood 94, 3737–3747 (1999).

  25. 25

    Davis, J.M. et al. Novel BTB/POZ domain zinc-finger protein, LRF, is a potential target of the LAZ-3/BCL-6 oncogene. Oncogene 18, 365–375 (1999).

  26. 26

    He, L.Z., Merghoub, T. & Pandolfi, P.P. In vivo analysis of the molecular pathogenesis of acute promyelocytic leukemia in the mouse and its therapeutic implications. Oncogene 18, 5278–5292 (1999).

  27. 27

    Bhatia, M. et al. Bone morphogenetic proteins regulate the developmental program of human hematopoietic stem cells. J. Exp. Med. 189, 1139–1148 (1999).

  28. 28

    Van Oostveen, J., Bijl, J., Raaphorst, F., Walboomers, J. & Meijer, C. The role of homeobox genes in normal hematopoiesis and hematological malignancies. Leukemia 13, 1675–1690 (1999).

  29. 29

    Tybulewicz, V.L., Crawford, C.E., Jackson, P.K., Bronson, R.T. & Mulligan, R.C. Neonatal lethality and lymphopenia in mice with a homozygous disruption of the c-abl proto-oncogene. Cell 65, 1153–1163 (1991).

  30. 30

    Ruthardt, M. et al. Opposite effects of the acute promyelocytic leukemia PML-retinoic acid receptor α (RAR α) and PLZF-RAR α fusion proteins on retinoic acid signalling. Mol. Cell. Biol. 17, 4859–4869 (1997).

  31. 31

    Kessel, M. & Gruss, P. Homeotic transformations of murine vertebrae and concomitant alteration of Hox codes by retinoic acid. Cell 67, 89–104 (1991).

  32. 32

    Wilkinson, D.G. Whole-mount in situ hybridization of vertebrate embryo. in In Situ hybridization: A Practical Approach (ed. Wilkinson, D.G.) 75–83 (IRL, Oxford, 1992).

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Acknowledgements

We thank D. Ruggero and S. Pizette for discussions; M. Branford and C. Kaduro for help with experiments; J.H. Dong and V. Soares for help with the generation and the management of the Zfp145 mutants; N. Lampin and B. Gumbiner for help and advice with the electron scanning microscopy analysis; D. Duboule and P. Gruss for Hoxd and Hoxa probes, respectively; M. Scott and A. Joyner for Ptch and Gli probes, respectively; M. Capecchi and A. Boulet for Hoxc6 and Hoxc8 probes; and P.G. Pelicci and M. Ruthardt for a PLZF antibody and probe. P.P.P. is a Scholar of the Leukemia and Lymphoma Society (formerly known as the Leukemia Society of America). L.N. is an Assistant Investigator of the Howard Hughes Medical Institute. This work was supported by the NIH (CA-08748 and CA-71692 awarded to Sloan-Kettering Institute and P.P.P.).

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Correspondence to Pier Paolo Pandolfi.

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