Over-expression of either MECP2-e1 or MECP2-e2 in neuronally differentiated cells results in different patterns of gene expression

Marija Orlic-Milacic, Liana Kaufman, Anna Mikhailov, Aaron Y.L. Cheung, Huda Mahmood, James Ellis, Peter J. Gianakopoulos, Berge A. Minassian, John B. Vincent

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9 Scopus citations

Abstract

Mutations in MECP2 are responsible for the majority of Rett syndrome cases. MECP2 is a regulator of transcription, and has two isoforms, MECP2-e1 and MECP2-e2. There is accumulating evidence that MECP2-e1 is the etiologically relevant variant for Rett. In this study we aim to detect genes that are differentially transcribed in neuronal cells over-expressing either of these two MECP2 isoforms. The human neuroblastoma cell line SK-N-SH was stably infected by lentiviral vectors overexpressing MECP2-e1, MECP2-e2, or eGFP, and were then differentiated into neurons. The same lentiviral constructs were also used to infect mouse Mecp2 knockout (Mecp2tm1.1Bird) fibroblasts. RNA from these cells was used for microarray gene expression analysis. For the human neuronal cells, ∼800 genes showed >three-fold change in expression level with the MECP2-e1 construct, and ,230 with MECP2-e2 (unpaired t-test, uncorrected p value <0.05). We used quantitative RT-PCR to verify microarray results for 41 of these genes. We found significant up-regulation of several genes resulting from overexpression of MECP2-e1 including SRPX2, NAV3, NPY1R, SYN3, and SEMA3D. DOCK8 was shown via microarray and qRT-PCR to be upregulated in both SK-N-SH cells and mouse fibroblasts. Both isoforms up-regulated GABRA2, KCNA1, FOXG1 and FOXP2. Down-regulation of expression in the presence of MECP2-e1 was seen with UNC5C and RPH3A. Understanding the biology of these differentially transcribed genes and their role in neurodevelopment may help us to understand the relative functions of the two MECP2 isoforms, and ultimately develop a better understanding of RTT etiology and determine the clinical relevance of isoform-specific mutations.

Original languageEnglish (US)
Article numbere91742
JournalPloS one
Volume9
Issue number4
DOIs
StatePublished - Apr 3 2014

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Orlic-Milacic, M., Kaufman, L., Mikhailov, A., Cheung, A. Y. L., Mahmood, H., Ellis, J., Gianakopoulos, P. J., Minassian, B. A., & Vincent, J. B. (2014). Over-expression of either MECP2-e1 or MECP2-e2 in neuronally differentiated cells results in different patterns of gene expression. PloS one, 9(4), [e91742]. https://doi.org/10.1371/journal.pone.0091742