Digital Object Identifier / DOI

https://doi.org/10.1007/978-1-62703-703-7_5

Department

Natural Sciences and Mathematics

Document Type

Article

Publication Date

1-1-2014

ISSN

1940-6029

Volume

1095

First Page

73

Last Page

86

Abstract

In animals, the Microprocessor complex cleaves primary transcripts of microRNAs (pri-miRNAs) to produce precursor microRNAs in the nucleus. The core components of Microprocessor include the Drosha ribonuclease and its RNA-binding partner protein DiGeorge critical region 8 (DGCR8). DGCR8 has been shown to tightly bind an Fe(III) heme cofactor, which activates its pri-miRNA processing activity. Here we describe how to reconstitute pri-miRNA processing using recombinant human Drosha and DGCR8 proteins. In particular, we present the procedures for expressing and purifying DGCR8 as an Fe(III) heme-bound dimer, the most active form of this protein, and for estimating its heme content.

PubMed ID

24166303

Included in

Biochemistry Commons

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