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Status |
Public on Dec 17, 2012 |
Title |
Expression profiling in primary embryonic fibroblasts deficient for the N-terminal truncation of Lats1 |
Organism |
Mus musculus |
Experiment type |
Expression profiling by array
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Summary |
Lats1 and Lats2 are the mediators of the Hippo pathway that regulates tissue growth and proliferation. Lats1 and Lats2 kinases share 85% sequence identity in the kinase domain. However, their non-kinase regions at the N-terminus are distinct except for Lats conserved domain 1 (LCD1) and LCD2, suggesting that their N-terminal regions are important for Lats1/2-specific functions. In this study, we generated Lats1 knockout mice disrupting the N-terminal region containing LCD1 (Lats1ΔN/ΔN). We show that some Lats1ΔN/ΔN mice were born safely and grew normally. However, mouse embryonic fibroblasts (MEFs) from Lats1ΔN/ΔN mice displayed drastic defects in mitosis, showing enhanced centrosome overduplication, chromosomal misalignment, multipolar spindle formation, chromosome bridging, and cytokinesis failure. Moreover, they displayed accelerated cell cycle and cell growth bypassing a cell-cell contact inhibition like tumor cells, and exhibited anchorage independent growth. Indeed, Lats1ΔN/ΔN MEFs produced tumors in nude mice after subcutaneous injection, although the tumor growth rate was much slower than ordinary cancer cells. Furthermore, Yap, the key transcriptional co-activator in the Hippo pathway, was overexpressed and retained stable in Lats1ΔN/ΔN MEFs under high cell density, and expression of Lats2 mRNA were down-regulated.
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Overall design |
Total RNAs were extracted from MEFs under high or low cell density using miRNeasy extraction kit (Qiagen). A Dye-swapped experiment was performed by hybridizing complimentary RNA (cRNA) labeled with either Cyanine (Cy) -3 or Cy-5 (Perkin-Elmer) onto Whole Mouse Genome Oligo Microarray (G4122F; Agilent Technologies). Co-hybridizations were performed and data from Cy3 or Cy5 channels were analyzed as normalized signal intensities rather than as log ratios corresponding to each array.
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Contributor(s) |
Okuzaki D, Nojima H |
Citation(s) |
23230145 |
Submission date |
May 09, 2012 |
Last update date |
May 10, 2018 |
Contact name |
Daisuke Okuzaki |
E-mail(s) |
dokuzaki@biken.osaka-u.ac.jp
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Phone |
+81-6-6879-4935
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Organization name |
Osaka univ.
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Department |
Immunology Frontier Research Center
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Lab |
Human Immunology (Single Cell Genomics)
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Street address |
Yamadaoka 3-1
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City |
Suita |
State/province |
Osaka |
ZIP/Postal code |
565-0871 |
Country |
Japan |
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Platforms (1) |
GPL4134 |
Agilent-014868 Whole Mouse Genome Microarray 4x44K G4122F (Feature Number version) |
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Samples (8)
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Relations |
BioProject |
PRJNA164681 |