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DNA-binding domain of myelin-gene regulatory factor: purification, crystallization and X-ray analysis.


ABSTRACT: The myelin sheath, which envelops axons in the vertebrate central nervous system, is crucial for the rapid conduction of action potentials. Myelin-gene regulatory factor (MRF) is a recently identified transcription factor that is required for myelin-sheath formation. Loss of MRF leads to demyelinating diseases and motor learning deficiency. MRF is a membrane-bound transcription factor that undergoes autocleavage from the endoplasmic reticulum membrane. The N-terminus of MRF contains a DNA-binding domain (DBD) that functions as a homotrimer. In this study, the MRF DBD was cloned, purified and crystallized in order to understand the molecular mechanism that regulates the transcription of myelin genes. Selenomethionine was subsequently introduced into the crystals to obtain the phases for the MRF DBD structure. The native and selenomethionine-labelled crystals exhibited diffraction to 2.50 and 2.51?Å resolution, respectively. The crystals belonged to space group P321 and the selenomethionine-labelled crystals had unit-cell parameters a = 104.0, b = 104.0, c = 46.7?Å, ? = 90, ? = 90, ? = 120°. The calculated Matthews coefficient was 3.04?Å3 Da-1 and the solvent content was 59.5%, indicating the presence of one MRF DBD molecule in the asymmetric unit.

SUBMITTER: Wu W 

PROVIDER: S-EPMC5505243 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature

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DNA-binding domain of myelin-gene regulatory factor: purification, crystallization and X-ray analysis.

Wu WenYu W   Zhen Xiangkai X   Shi Ning N  

Acta crystallographica. Section F, Structural biology communications 20170617 Pt 7


The myelin sheath, which envelops axons in the vertebrate central nervous system, is crucial for the rapid conduction of action potentials. Myelin-gene regulatory factor (MRF) is a recently identified transcription factor that is required for myelin-sheath formation. Loss of MRF leads to demyelinating diseases and motor learning deficiency. MRF is a membrane-bound transcription factor that undergoes autocleavage from the endoplasmic reticulum membrane. The N-terminus of MRF contains a DNA-bindin  ...[more]

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