K-Ras Lys-42 is crucial for its signaling, cell migration, and invasion.

Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE

Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology

Cell Movement* ; MAP Kinase Signaling System*; Proto-Oncogene Proteins p21(ras)/*metabolism; Animals ; Checkpoint Kinase 2/genetics ; Checkpoint Kinase 2/metabolism ; Extracellular Signal-Regulated MAP Kinases/genetics ; Extracellular Signal-Regulated MAP Kinases/metabolism ; Flavones/pharmacology ; Focal Adhesion Kinase 1/genetics ; Focal Adhesion Kinase 1/metabolism ; HEK293 Cells ; Humans ; MCF-7 Cells ; Mice ; NIH 3T3 Cells ; Neoplasm Invasiveness/genetics ; Neoplasm Invasiveness/pathology ; Proto-Oncogene Proteins p21(ras)/genetics ; Sumoylation/drug effects ; Sumoylation/genetics ; Ubiquitin-Conjugating Enzymes/antagonists & inhibitors ; Ubiquitin-Conjugating Enzymes/genetics ; Ubiquitin-Conjugating Enzymes/metabolism ; Zinc Finger E-box-Binding Homeobox 1/genetics ; Zinc Finger E-box-Binding Homeobox 1/metabolism ; Zonula Occludens-1 Protein/genetics ; Zonula Occludens-1 Protein/metabolism

Ras proteins participate in multiple signal cascades, regulating crucial cellular processes, including cell survival, proliferation, and differentiation. We have previously reported that Ras proteins are modified by sumoylation and that Lys-42 plays an important role in mediating the modification. In the current study, we further investigated the role of Lys-42 in regulating cellular activities of K-Ras. Inducible expression of K-Ras ^V12 led to the activation of downstream components, including c-RAF, MEK1, and extracellular signal-regulated kinases (ERKs), whereas expression of K-Ras ^V12/R42 mutant compromised the activation of the RAF/MEK/ERK signaling axis. Expression of K-Ras ^V12/R42 also led to reduced phosphorylation of several other protein kinases, including c-Jun N-terminal kinase (JNK), Chk2, and focal adhesion kinase (FAK). Significantly, K-Ras ^V12/R42 expression inhibited cellular migration and invasion in vitro in multiple cell lines, including transformed pancreatic cells. Given that K-Ras plays a crucial role in mediating oncogenesis in the pancreas, we treated transformed pancreatic cells of both BxPC-3 and MiaPaCa-2 with 2-D08, a small ubiquitin-like modifier (SUMO) E2 inhibitor. Treatment with the compound inhibited cell migration in a concentration-dependent manner, which was correlated with a reduced level of K-Ras sumoylation. Moreover, 2-D08 suppressed expression of ZEB1 (a mesenchymal cell marker) with concomitant induction of ZO-1 (an epithelial cell marker). Combined, our studies strongly suggest that posttranslational modification(s), including sumoylation mediated by Lys-42, plays a crucial role in K-Ras activities in vivo .
(© 2018 Choi et al.)

Cell. 1997 May 2;89(3):457-67. (PMID: 9150145)
Cancer Cell. 2015 Aug 10;28(2):170-82. (PMID: 26267534)
Proc Natl Acad Sci U S A. 2013 Dec 17;110(51):20593-8. (PMID: 24297914)
Nat Med. 2013 Aug;19(8):1047-53. (PMID: 23817022)
Proc Natl Acad Sci U S A. 2015 Jan 20;112(3):779-84. (PMID: 25561545)
Nature. 2006 May 25;441(7092):424-30. (PMID: 16724053)
Mol Membr Biol. 2009 Jan;26(1):80-92. (PMID: 19115142)
Cell. 2007 Jan 26;128(2):233-4. (PMID: 17254959)
Carcinogenesis. 2011 Sep;32(9):1299-304. (PMID: 21665887)
Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):E1724-33. (PMID: 25805818)
BMC Cancer. 2016 Mar 18;16:236. (PMID: 26988558)
Oncotarget. 2014 Sep 30;5(18):7988-8013. (PMID: 25237759)
Expert Rev Anticancer Ther. 2013 Mar;13(3):327-37. (PMID: 23477519)
Clin Cancer Res. 2015 Apr 15;21(8):1819-27. (PMID: 25878363)
Cell. 1998 Nov 13;95(4):447-50. (PMID: 9827797)
Nat Rev Mol Cell Biol. 2011 Dec 22;13(1):39-51. (PMID: 22189424)
J Clin Invest. 2007 May;117(5):1223-5. (PMID: 17476354)
Mol Cell. 2006 Feb 17;21(4):481-93. (PMID: 16483930)
Science. 1990 Aug 10;249(4969):635-40. (PMID: 2116664)
Mol Cell. 2006 Mar 3;21(5):679-87. (PMID: 16507365)
PLoS One. 2012;7(7):e40435. (PMID: 22848379)
J Cell Physiol. 2015 Mar;230(3):610-9. (PMID: 25158650)
Nature. 1993 Dec 16;366(6456):643-54. (PMID: 8259209)
Cell. 2016 Apr 21;165(3):643-55. (PMID: 27104980)
Cell. 2000 Oct 13;103(2):227-38. (PMID: 11057896)
Annu Rev Immunol. 2006;24:771-800. (PMID: 16551266)
Oncotarget. 2017 Dec 15;9(4):4440-4450. (PMID: 29435114)
Curr Biol. 2010 Aug 10;20(15):1372-7. (PMID: 20655225)
Proc Natl Acad Sci U S A. 2012 Jul 3;109(27):10843-8. (PMID: 22711838)
Bioorg Med Chem Lett. 2014 Feb 15;24(4):1094-7. (PMID: 24468414)

R01 CA116034 United States CA NCI NIH HHS; R01 GM086171 United States GM NIGMS NIH HHS; P30 ES000260 United States ES NIEHS NIH HHS; R01 CA216987 United States CA NCI NIH HHS; R01 CA265410 United States CA NCI NIH HHS

Keywords: K-Ras; Ras protein; cell invasion; cell migration; domain structure; lysine residue; posttranslational modification (PTM); signaling; sumoylation; transformation

0 (2',3',4'-trihydroxyflavone)
0 (Flavones)
0 (KRAS protein, human)
0 (TJP1 protein, human)
0 (Tjp1 protein, mouse)
0 (ZEB1 protein, human)
0 (ZEB1 protein, mouse)
0 (Zinc Finger E-box-Binding Homeobox 1)
0 (Zonula Occludens-1 Protein)
EC 2.3.2.23 (Ubiquitin-Conjugating Enzymes)
EC 2.7.1.11 (Checkpoint Kinase 2)
EC 2.7.10.2 (Focal Adhesion Kinase 1)
EC 2.7.10.2 (PTK2 protein, human)
EC 2.7.10.2 (Ptk2 protein, mouse)
EC 2.7.11.1 (CHEK2 protein, human)
EC 2.7.11.1 (Chek2 protein, mouse)
EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases)
EC 3.6.5.2 (Hras protein, mouse)
EC 3.6.5.2 (Proto-Oncogene Proteins p21(ras))

Date Created: 20180920 Date Completed: 20190319 Latest Revision: 20230309

20230309

PMC6231119

10.1074/jbc.RA118.003723

30228186