Skip to main content
SpringerLink
Log in

Alternate PAX3 and PAX7 C-terminal isoforms in myogenic differentiation and sarcomagenesis

  • Research Articles
  • Published:
Clinical and Translational Oncology Aims and scope Submit manuscript

Abstract

Objective

Pax3 and Pax7 are closely related genes that are involved in commitment of cells to a myogenic lineage during skeletal muscle development and regeneration. Several Pax3 and Pax7 transcripts are expressed from the genes, generating different isoforms with potentially distinct DNA binding and transactivation properties. The aim of this study was to investigate the implication of Pax3 and Pax7 C-terminal isoforms during myogenic differentiation and tumorigenesis, since fusions involving these genes are commonly associated with alveolar rhabdomyosarcoma (ARMS).

Methods

Uncommitted (mouse mesenchymal stem cells, MSCs) and committed (C2C12) myogenic precursor cells were stably transfected with PAX3/FKHR and PAXC7/FKHR fusion genes. We analysed gene and protein expression comparing the newly generated cells with the parental cells, to determine the functional importance of Pax3 and Pax7 C-terminal isoforms.

Results

We found that the transcript Pax3c was expressed at low levels in undifferentiated C2C12 and MSCs cells, but its expression levels increased considerably at later stages of differentiation. However, expression levels of Pax3d transcript increased only slightly after differentiation. Pax7 transcripts, present before differentiation in committed C2C12 cells, but absent in uncommitted MSCs, increased noticeably in MSCs after differentiation. We also found that the presence of PAX/FKHR fusions prevented both C2C12 and MSC cells from terminal myogenic differentiation and increased the expression of discrete endogenous Pax3/7 transcripts, in particular Pax3d and Pax7B. Conclusions Our results suggest that both Pax3 and Pax7 transcripts are required for commitment of cells to the myogenic lineage, with each transcript having a distinct role. More specifically, the Pax3c isoform may be required for terminal myogenic differentiation whereas the Pax3d isoform may be involved in undifferentiated cell maintenance and/or proliferation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Chi N, Epstein JA (2002) Getting your Pax straight: Pax proteins in development and disease. Trends Genet 18:41–47

    Article  PubMed  CAS  Google Scholar 

  2. Mikkola I, Bruun JA, Bjorkoy G et al (1999) Phosphorylation of the transactivation domain of Pax6 by extracellular signal-regulated kinase and p38 mitogen-activated protein kinase. J Biol Chem 274:15115–15126

    Article  PubMed  CAS  Google Scholar 

  3. Underhill DA, Gros P (1997) The paired-domain regulates DNA binding by the homeodomain within the intact Pax-3 protein. J Biol Chem 272:14175–14182

    Article  PubMed  CAS  Google Scholar 

  4. Lamey TM, Koenders A, Ziman M (2004) Pax genes in myogenesis: alternate transcripts add complexity. Histol Histopathol 19:1289–1300

    PubMed  CAS  Google Scholar 

  5. Vogan KJ, Underhill DA, Gros P (1996) An alternative splicing event in the Pax-3 paired domain identifies the linker region as a key determinant of paired domain DNA-binding activity. Mol Cell Biol 16:6677–6686

    PubMed  CAS  Google Scholar 

  6. Ziman MR, Fletcher S, Kay PH (1997) Alternate Pax7 transcripts are expressed specifically in skeletal muscle, brain and other organs of adult mice. Int J Biochem Cell Biol 29:1029–1036

    Article  PubMed  CAS  Google Scholar 

  7. White RB, Ziman MR (2008) Genome-wide discovery of Pax7 target genes during development. Physiol Genomics 33:41–49

    Article  PubMed  CAS  Google Scholar 

  8. Du S, Lawrence EJ, Strzelecki D et al (2005) Coexpression of alternatively spliced forms of PAX3, PAX7, PAX3-FKHR and PAX7-FKHR with distinct DNA binding and transactivation properties in rhabdomyosarcoma. Int J Cancer 115:85–92

    Article  PubMed  CAS  Google Scholar 

  9. Barr FG, Fitzgerald JC, Ginsberg JP et al (1999) Predominant expression of alternative PAX3 and PAX7 forms in myogenic and neural tumor cell lines. Cancer Res 59:5443–5448

    PubMed  CAS  Google Scholar 

  10. Pritchard C, Grosveld G, Hollenbach AD (2003) Alternative splicing of Pax3 produces a transcriptionally inactive protein. Gene 305:61–69

    Article  PubMed  CAS  Google Scholar 

  11. Parker CJ, Shawcross SG, Li H et al (2004) Expression of PAX 3 alternatively spliced transcripts and identification of two new isoforms in human tumors of neural crest origin. Int J Cancer 108:314–320

    Article  PubMed  CAS  Google Scholar 

  12. Wang Q, Kumar S, Slevin M et al (2006) Functional analysis of alternative isoforms of the transcription factor PAX3 in melanocytes in vitro. Cancer Res 66:8574–8580

    Article  PubMed  CAS  Google Scholar 

  13. Barber TD, Barber MC, Cloutier TE et al (1999) PAX3 gene structure, alternative splicing and evolution. Gene 237:311–319

    Article  PubMed  CAS  Google Scholar 

  14. Yaffe D, Saxel O (1977) A myogenic cell line with altered serum requirements for differentiation. Differentiation 7:159–166

    Article  PubMed  CAS  Google Scholar 

  15. Wakitani S, Saito T, Caplan AI (1995) Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve 18:1417–1426

    Article  PubMed  CAS  Google Scholar 

  16. Pittenger M, Vanguri P, Simonetti D et al (2002) Adult mesenchymal stem cells: potential for muscle and tendon regeneration and use in gene therapy. J Musculoskelet Neuronal Interact 2:309–320

    PubMed  CAS  Google Scholar 

  17. Charytonowicz E, Cordon-Cardo C, Matushansky I et al (2009) Alveolar rhabdomyosarcoma: is the cell of origin a mesenchymal stem cell? Cancer Lett 279:126–136

    Article  PubMed  CAS  Google Scholar 

  18. Gang EJ, Bosnakovski D, Simsek T et al (2008) Pax3 activation promotes the differentiation of mesenchymal stem cells toward the myogenic lineage. Exp Cell Res 314:1721–1733

    Article  PubMed  CAS  Google Scholar 

  19. Wannenes F, Caprio M, Gatta L et al (2008) Androgen receptor expression during C2C12 skeletal muscle cell line differentiation. Mol Cell Endocrinol 292:11–19

    Article  PubMed  CAS  Google Scholar 

  20. Blake JA, Ziman MR (2005) Pax3 transcripts in melanoblast development. Dev Growth Differ 47:627–635

    Article  PubMed  CAS  Google Scholar 

  21. de Alava E, Ladanyi M, Rosai J et al (1995) Detection of chimeric transcripts in desmoplastic small round cell tumor and related developmental tumors by reverse transcriptase polymerase chain reaction. A specific diagnostic assay. Am J Pathol 147:1584–1591

    PubMed  Google Scholar 

  22. Graf Finckenstein F, Shahbazian V, Davicioni E et al (2008) PAX-FKHR function as pangenes by simultaneously inducing and inhibiting myogenesis. Oncogene 27:2004–2014

    Article  PubMed  CAS  Google Scholar 

  23. Czerny T, Schaffner G, Busslinger M (1993) DNA sequence recognition by Pax proteins: bipartite structure of the paired domain and its binding site. Genes Dev 7:2048–2061

    Article  PubMed  CAS  Google Scholar 

  24. Vogan KJ, Gros P (1997) The C-terminal subdomain makes an important contribution to the DNA binding activity of the Pax-3 paired domain. J Biol Chem 272:28289–28295

    Article  PubMed  CAS  Google Scholar 

  25. Wang Q, Fang WH, Krupinski J et al (2008) Pax genes in embryogenesis and oncogenesis. J Cell Mol Med 12:2281–2294

    Article  PubMed  CAS  Google Scholar 

  26. Deneen B, Denny CT (2001) Loss of p16 pathways stabilizes EWS/FLI1 expression and complements EWS/FLI1 mediated transformation. Oncogene 20:6731–6741

    Article  PubMed  CAS  Google Scholar 

  27. Xia SJ, Barr FG (2004) Analysis of the transforming and growth suppressive activities of the PAX3-FKHR oncoprotein. Oncogene 23:6864–6871

    Article  PubMed  CAS  Google Scholar 

  28. Vorobyov E, Horst J (2004) Expression of two protein isoforms of PAX7 is controlled by competing cleavage-polyadenylation and splicing. Gene 342:107–112

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Exercise Biomedical and Health Science, Edith Cowan University, 270 Joondalup Drive, Joondalup, WA, 6027, Australia

    Elizabeth Charytonowicz & Mel Ziman

  2. Irving Cancer Research Center, Columbia University, New York, USA

    Elizabeth Charytonowicz, Igor Matushansky, Mireia Castillo-Martin, Todd Hricik & Carlos Cordon-Cardo

  3. School of Pathology and Laboratory Medicine, University of Western Australia, Crawley, Australia

    Mel Ziman

Authors
  1. Elizabeth Charytonowicz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Igor Matushansky
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mireia Castillo-Martin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Todd Hricik
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Carlos Cordon-Cardo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mel Ziman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mel Ziman.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Charytonowicz, E., Matushansky, I., Castillo-Martin, M. et al. Alternate PAX3 and PAX7 C-terminal isoforms in myogenic differentiation and sarcomagenesis. Clin Transl Oncol 13, 194–203 (2011). https://doi.org/10.1007/s12094-011-0640-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12094-011-0640-y

Keywords

Search

Navigation