Der Lab Collaborator Featured on Cover of Genes & Cancer

GenesandCancer_LaurenHuffLauren Huff, a member of Dr. Adrienne Cox’s lab at UNC Lineberger and frequent collaborator of the Der lab, is featured on the latest issue of Genes & Cancer (Nov/Dec 2013). Lauren successfully defended her thesis November 4, 2013. She is the first author of the following article.

The Role of Ect2 Nuclear RhoGEF Activity in Ovarian Cancer Cell Transformation

  1. Lauren P. Huff1
  2. Molly J. DeCristo2
  3. Dimitri Trembath3
  4. Pei Fen Kuan4
  5. Margaret Yim5
  6. Jinsong Liu6
  7. Danielle R. Cook7
  8. C. Ryan Miller3,8
  9. Channing J. Der1,8
  10. Adrienne D. Cox1,8,9
  1. 1Department of Pharmacology, University of North Carolina, Chapel Hill, NC, USA
  2. 2Department of Biology, University of North Carolina, Chapel Hill, NC, USA
  3. 3Department of Pathology & Laboratory Medicine, University of North Carolina, Chapel Hill, NC, USA
  4. 4Department of Biostatistics, University of North Carolina, Chapel Hill, NC, USA
  5. 5Department of Molecular Biophysics & Biochemistry, Yale University, New Haven, CT, USA
  6. 6Department of Pathology, MD Anderson Cancer Center, Houston, TX, USA
  7. 7School of Pharmacy, University of North Carolina, Chapel Hill, NC, USA
  8. 8Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, USA
  9. 9Department of Radiation Oncology, University of North Carolina, Chapel Hill, NC, USA
  1. Adrienne D. Cox, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7295, USA (Email: adrienne_cox@med.unc.edu).

Abstract

Ect2, a Rho guanine nucleotide exchange factor (RhoGEF), is atypical among RhoGEFs in its predominantly nuclear localization in interphase cells. One current model suggests that Ect2 mislocalization drives cellular transformation by promoting aberrant activation of cytoplasmic Rho family GTPase substrates. However, in ovarian cancers, where Ect2 is both amplified and overexpressed at the mRNA level, we observed that the protein is highly expressed and predominantly nuclear and that nuclear but not cytoplasmic Ect2 increases with advanced disease. Knockdown of Ect2 in ovarian cancer cell lines impaired their anchorage-independent growth without affecting their growth on plastic. Restoration of Ect2 expression rescued the anchorage-independent growth defect, but not if either the DH catalytic domain or the nuclear localization sequences of Ect2 were mutated. These results suggested a novel mechanism whereby Ect2 could drive transformation in ovarian cancer cells by acting as a RhoGEF specifically within the nucleus. Interestingly, Ect2 had an intrinsically distinct GTPase specificity profile in the nucleus versus the cytoplasm. Nuclear Ect2 bound preferentially to Rac1, while cytoplasmic Ect2 bound to RhoA but not Rac. Consistent with nuclear activation of endogenous Rac, Ect2 overexpression was sufficient to recruit Rac effectors to the nucleus, a process that required a functional Ect2 catalytic domain. Furthermore, expression of active nuclearly targeted Rac1 rescued the defect in transformed growth caused by Ect2 knockdown. Our work suggests a novel mechanism of Ect2-driven transformation, identifies subcellular localization as a regulator of GEF specificity, and implicates activation of nuclear Rac1 in cellular transformation.

Advertisements

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s