Mechanism for the alteration of the substrate specificities of template-independent RNA polymerases

Yukimatsu Toh, Daijiro Takeshita, Takashi Nagaike, Tomoyuki Numata, Kozo Tomita

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

PolyA polymerase (PAP) adds a polyA tail onto the 3′-end of RNAs without a nucleic acid template, using adenosine-5′-triphosphate (ATP) as a substrate. The mechanism for the substrate selection by eubacterial PAP remains obscure. Structural and biochemical studies of Escherichia coli PAP (EcPAP) revealed that the shape and size of the nucleobase-interacting pocket of EcPAP are maintained by an intra-molecular hydrogen-network, making it suitable for the accommodation of only ATP, using a single amino acid, Arg 197. The pocket structure is sustained by interactions between the catalytic domain and the RNA-binding domain. EcPAP has a flexible basic C-terminal region that contributes to optimal RNA translocation for processive adenosine 5′-monophosphate (AMP) incorporations onto the 3′-end of RNAs. A comparison of the EcPAP structure with those of other template-independent RNA polymerases suggests that structural changes of domain(s) outside the conserved catalytic core domain altered the substrate specificities of the template-independent RNA polymerases.

Original languageEnglish
Pages (from-to)232-243
Number of pages12
JournalStructure
Volume19
Issue number2
DOIs
Publication statusPublished - Feb 9 2011

Fingerprint

DNA-Directed RNA Polymerases
Substrate Specificity
Escherichia coli
Catalytic Domain
RNA
Adenosine Triphosphate
Catalytic RNA
Adenosine Monophosphate
Nucleic Acids
Hydrogen
Amino Acids

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Cite this

Mechanism for the alteration of the substrate specificities of template-independent RNA polymerases. / Toh, Yukimatsu; Takeshita, Daijiro; Nagaike, Takashi; Numata, Tomoyuki; Tomita, Kozo.

In: Structure, Vol. 19, No. 2, 09.02.2011, p. 232-243.

Research output: Contribution to journalArticle

Toh, Yukimatsu ; Takeshita, Daijiro ; Nagaike, Takashi ; Numata, Tomoyuki ; Tomita, Kozo. / Mechanism for the alteration of the substrate specificities of template-independent RNA polymerases. In: Structure. 2011 ; Vol. 19, No. 2. pp. 232-243.
@article{3077e3cf5a5b436d8b251331929f657a,
title = "Mechanism for the alteration of the substrate specificities of template-independent RNA polymerases",
abstract = "PolyA polymerase (PAP) adds a polyA tail onto the 3′-end of RNAs without a nucleic acid template, using adenosine-5′-triphosphate (ATP) as a substrate. The mechanism for the substrate selection by eubacterial PAP remains obscure. Structural and biochemical studies of Escherichia coli PAP (EcPAP) revealed that the shape and size of the nucleobase-interacting pocket of EcPAP are maintained by an intra-molecular hydrogen-network, making it suitable for the accommodation of only ATP, using a single amino acid, Arg 197. The pocket structure is sustained by interactions between the catalytic domain and the RNA-binding domain. EcPAP has a flexible basic C-terminal region that contributes to optimal RNA translocation for processive adenosine 5′-monophosphate (AMP) incorporations onto the 3′-end of RNAs. A comparison of the EcPAP structure with those of other template-independent RNA polymerases suggests that structural changes of domain(s) outside the conserved catalytic core domain altered the substrate specificities of the template-independent RNA polymerases.",
author = "Yukimatsu Toh and Daijiro Takeshita and Takashi Nagaike and Tomoyuki Numata and Kozo Tomita",
year = "2011",
month = "2",
day = "9",
doi = "10.1016/j.str.2010.12.006",
language = "English",
volume = "19",
pages = "232--243",
journal = "Structure with Folding & design",
issn = "0969-2126",
publisher = "Cell Press",
number = "2",

}

TY - JOUR

T1 - Mechanism for the alteration of the substrate specificities of template-independent RNA polymerases

AU - Toh, Yukimatsu

AU - Takeshita, Daijiro

AU - Nagaike, Takashi

AU - Numata, Tomoyuki

AU - Tomita, Kozo

PY - 2011/2/9

Y1 - 2011/2/9

N2 - PolyA polymerase (PAP) adds a polyA tail onto the 3′-end of RNAs without a nucleic acid template, using adenosine-5′-triphosphate (ATP) as a substrate. The mechanism for the substrate selection by eubacterial PAP remains obscure. Structural and biochemical studies of Escherichia coli PAP (EcPAP) revealed that the shape and size of the nucleobase-interacting pocket of EcPAP are maintained by an intra-molecular hydrogen-network, making it suitable for the accommodation of only ATP, using a single amino acid, Arg 197. The pocket structure is sustained by interactions between the catalytic domain and the RNA-binding domain. EcPAP has a flexible basic C-terminal region that contributes to optimal RNA translocation for processive adenosine 5′-monophosphate (AMP) incorporations onto the 3′-end of RNAs. A comparison of the EcPAP structure with those of other template-independent RNA polymerases suggests that structural changes of domain(s) outside the conserved catalytic core domain altered the substrate specificities of the template-independent RNA polymerases.

AB - PolyA polymerase (PAP) adds a polyA tail onto the 3′-end of RNAs without a nucleic acid template, using adenosine-5′-triphosphate (ATP) as a substrate. The mechanism for the substrate selection by eubacterial PAP remains obscure. Structural and biochemical studies of Escherichia coli PAP (EcPAP) revealed that the shape and size of the nucleobase-interacting pocket of EcPAP are maintained by an intra-molecular hydrogen-network, making it suitable for the accommodation of only ATP, using a single amino acid, Arg 197. The pocket structure is sustained by interactions between the catalytic domain and the RNA-binding domain. EcPAP has a flexible basic C-terminal region that contributes to optimal RNA translocation for processive adenosine 5′-monophosphate (AMP) incorporations onto the 3′-end of RNAs. A comparison of the EcPAP structure with those of other template-independent RNA polymerases suggests that structural changes of domain(s) outside the conserved catalytic core domain altered the substrate specificities of the template-independent RNA polymerases.

UR - http://www.scopus.com/inward/record.url?scp=79551705393&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79551705393&partnerID=8YFLogxK

U2 - 10.1016/j.str.2010.12.006

DO - 10.1016/j.str.2010.12.006

M3 - Article

C2 - 21300291

AN - SCOPUS:79551705393

VL - 19

SP - 232

EP - 243

JO - Structure with Folding & design

JF - Structure with Folding & design

SN - 0969-2126

IS - 2

ER -