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Literature DB >> 26253968

Preparation of Human Nuclear RNA m⁶A Methyltransferases and Demethylases and Biochemical Characterization of Their Catalytic Activity.

Abstract

N(6)-Methyladenosine (m(6)A) represents the most prevalent internal modification in messenger and long noncoding RNAs. There has been a surge of interest toward understanding the biological significance of m(6)A modification. In this chapter, we describe the methods for biochemically studying the recently uncovered m(6)A methyltransferases (METTL3 and METTL14) and demethylases (FTO and ALKBH5). How to express these proteins, perform their biochemistry reactions against various RNA probes, and characterize the methylation and demethylation activity will be discussed.

Entities: CellLine Chemical Disease Gene Species

Keywords: ALKBH5; Demethylation activity; FTO; METTL3–METTL14; Methylation activity; N(6)-methyladenosine; m(6)A demethylase; m(6)A methyltransferase

Mesh：

Substances：

Year: 2015 PMID： 26253968 PMCID： PMC5971108 DOI： 10.1016/bs.mie.2015.03.013

Source DB: PubMed Journal: Methods Enzymol ISSN： 0076-6879 Impact factor: 1.600

23 in total

1. Influenza viral mRNA contains internal N6-methyladenosine and 5'-terminal 7-methylguanosine in cap structures.

Authors: R M Krug; M A Morgan; A J Shatkin
Journal: J Virol Date: 1976-10 Impact factor: 5.103

2. 5'-Terminal and internal methylated nucleotide sequences in HeLa cell mRNA.

Authors: C M Wei; A Gershowitz; B Moss
Journal: Biochemistry Date: 1976-01-27 Impact factor: 3.162

3. Localization of N6-methyladenosine in the Rous sarcoma virus genome.

Authors: K Beemon; J Keith
Journal: J Mol Biol Date: 1977-06-15 Impact factor: 5.469

4. m(6)A RNA modification controls cell fate transition in mammalian embryonic stem cells.

Authors: Pedro J Batista; Benoit Molinie; Jinkai Wang; Kun Qu; Jiajing Zhang; Lingjie Li; Donna M Bouley; Ernesto Lujan; Bahareh Haddad; Kaveh Daneshvar; Ava C Carter; Ryan A Flynn; Chan Zhou; Kok-Seong Lim; Peter Dedon; Marius Wernig; Alan C Mullen; Yi Xing; Cosmas C Giallourakis; Howard Y Chang
Journal: Cell Stem Cell Date: 2014-10-16 Impact factor: 24.633

5. Comprehensive analysis of mRNA methylation reveals enrichment in 3' UTRs and near stop codons.

Authors: Kate D Meyer; Yogesh Saletore; Paul Zumbo; Olivier Elemento; Christopher E Mason; Samie R Jaffrey
Journal: Cell Date: 2012-05-17 Impact factor: 41.582

6. Induction of sporulation in Saccharomyces cerevisiae leads to the formation of N6-methyladenosine in mRNA: a potential mechanism for the activity of the IME4 gene.

Authors: Mary J Clancy; Mary Eileen Shambaugh; Candace S Timpte; Joseph A Bokar
Journal: Nucleic Acids Res Date: 2002-10-15 Impact factor: 16.971

7. Mapping of N6-methyladenosine residues in bovine prolactin mRNA.

Authors: S Horowitz; A Horowitz; T W Nilsen; T W Munns; F M Rottman
Journal: Proc Natl Acad Sci U S A Date: 1984-09 Impact factor: 11.205

8. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility.

Authors: Guanqun Zheng; John Arne Dahl; Yamei Niu; Peter Fedorcsak; Chun-Min Huang; Charles J Li; Cathrine B Vågbø; Yue Shi; Wen-Ling Wang; Shu-Hui Song; Zhike Lu; Ralph P G Bosmans; Qing Dai; Ya-Juan Hao; Xin Yang; Wen-Ming Zhao; Wei-Min Tong; Xiu-Jie Wang; Florian Bogdan; Kari Furu; Ye Fu; Guifang Jia; Xu Zhao; Jun Liu; Hans E Krokan; Arne Klungland; Yun-Gui Yang; Chuan He
Journal: Mol Cell Date: 2012-11-21 Impact factor: 17.970

Preparation of Human Nuclear RNA m⁶A Methyltransferases and Demethylases and Biochemical Characterization of Their Catalytic Activity.

1. Influenza viral mRNA contains internal N6-methyladenosine and 5'-terminal 7-methylguanosine in cap structures.

2. 5'-Terminal and internal methylated nucleotide sequences in HeLa cell mRNA.

3. Localization of N6-methyladenosine in the Rous sarcoma virus genome.

4. m(6)A RNA modification controls cell fate transition in mammalian embryonic stem cells.

5. Comprehensive analysis of mRNA methylation reveals enrichment in 3' UTRs and near stop codons.

6. Induction of sporulation in Saccharomyces cerevisiae leads to the formation of N6-methyladenosine in mRNA: a potential mechanism for the activity of the IME4 gene.

7. Mapping of N6-methyladenosine residues in bovine prolactin mRNA.

8. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility.

9. N6-methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO.

10. N6-methyladenosine-dependent regulation of messenger RNA stability.

1. Structural Basis for Cooperative Function of Mettl3 and Mettl14 Methyltransferases.

2. HIV Replication Is Increased by RNA Methylation METTL3/METTL14/WTAP Complex Activators.