Characterization of ReNCell for studying chromatin associated proteins MeCP2 and histone H1

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dc.contributor.author Kim, Bo Hyun "Cindy"
dc.date.accessioned 2022-08-05T18:51:28Z
dc.date.available 2022-08-05T18:51:28Z
dc.date.copyright 2022 en_US
dc.date.issued 2022-08-05
dc.identifier.uri http://hdl.handle.net/1828/14085
dc.description.abstract Methyl-CpG binding protein 2 (MeCP2) and histone H1 are important chromatin associated proteins. Both exhibit their own extent of complexity as MeCP2 is an intrinsically disordered protein (IDP) that interacts with many different partners involved in several cellular processes and histone H1 consists of 11 different subtypes each of them associated with different posttranslational modifications (PTMs). An interesting avenue for the study of these proteins is in neurons where MeCP2 is very abundant and histone H1 level is half that observed in other somatic tissues. Several reports in the past have proposed that this lower level of histone H1 is due to the abundance of MeCP2 which displaces histone H1. However, this hypothesis has been debated and there is no clear consensus. In an attempt to study this controversy, a cell model system ReNCell WT and MeCP2-KO was used that can be induced to differentiate into neurons. The protein levels, transcript levels and localization of histone H1 subtypes in these cells were analyzed using HPLC, RT-qPCR and immunofluorescence, respectively. The results show that ReNCell WT and MeCP2-KO do not exhibit significant differences in their relative amount of histone H1 protein and transcript level neither at the proliferative nor at the later differentiated stages. However, HPLC analyses show that the histone H1 subtypes of these two cell types exhibit significant elution differences probably resulting from differences in their PTM content. Immunofluorescence analyses show that WT ReNCell differentiation as determined by extension of dendritic or axonic processes can be seen to occur over the course of one week and there is a significant difference in the nuclear area of these two cells at 8 DIV. This study provides important preliminary data for future research in MeCP2 and histone H1 using this cell model system and show that MeCP2 may have a bearing on histone H1 PTMs. en_US
dc.language English eng
dc.language.iso en en_US
dc.rights Available to the World Wide Web en_US
dc.subject ReNCell en_US
dc.subject MeCP2 en_US
dc.subject Histone H1 en_US
dc.subject Epigenetics en_US
dc.title Characterization of ReNCell for studying chromatin associated proteins MeCP2 and histone H1 en_US
dc.type Thesis en_US
dc.contributor.supervisor Ausió, Juan
dc.degree.department Department of Biochemistry and Microbiology en_US
dc.degree.level Master of Science M.Sc. en_US
dc.description.scholarlevel Graduate en_US

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