Cooperative and anticooperative binding to a ribozyme

Philip C. Bevilacqua, Kenneth A. Johnson, Douglas H. Turner

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

The effects of guanosine 5′-monophosphate and 2′-deoxyguanosine 5′-monophosphate on the thermodynamics and kinetics of pyrene-labeled 5′ exon mimic (pyCUCU) binding to the catalytic RNA (ribozyme) from Tetrahymena thermophila have been determined by fluorescence titration and kinetics experiments at 15°C. pyCUCU binding to L-21 Sca I-truncated ribozyme is weaker by a factor of 5 in the presence of saturating guanosine 5′-monophosphate, whereas it is 4-fold stronger in the presence of saturating 2′-deoxyguanosine 5′-monophosphate. Results from kinetics experiments suggest that ant kooperative effects in the presence of guanosine 5′-monophosphate arise primarily from slower formation of tertiary contacts between the catalytic core of the ribozyme and the P1 duplex formed by pyCUCU and GGAGGG of the ribozyme. Conversely, cooperative effects in the presence of 2′-deoxyguanosine 5′-monophosphate arise primarily from slower disruption of tertiary contacts between the catalytic core of the ribozyme and the P1 duplex. Additional experiments suggest that these cooperative and anticooperative effects are not a function of the pyrene label, are not caused by a salt effect, and are not specific to one renaturation procedure for the ribozyme.

Original languageEnglish (US)
Pages (from-to)8357-8361
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number18
DOIs
StatePublished - Sep 15 1993

Fingerprint

Catalytic RNA
Guanosine Monophosphate
Catalytic Domain
Tetrahymena thermophila
Ants
Thermodynamics
Exons
Salts
Fluorescence
2'-deoxyguanosine 5'-phosphate

All Science Journal Classification (ASJC) codes

  • General

Cite this

@article{c8b1e4877c3c45839e569aef71ab9f56,
title = "Cooperative and anticooperative binding to a ribozyme",
abstract = "The effects of guanosine 5′-monophosphate and 2′-deoxyguanosine 5′-monophosphate on the thermodynamics and kinetics of pyrene-labeled 5′ exon mimic (pyCUCU) binding to the catalytic RNA (ribozyme) from Tetrahymena thermophila have been determined by fluorescence titration and kinetics experiments at 15°C. pyCUCU binding to L-21 Sca I-truncated ribozyme is weaker by a factor of 5 in the presence of saturating guanosine 5′-monophosphate, whereas it is 4-fold stronger in the presence of saturating 2′-deoxyguanosine 5′-monophosphate. Results from kinetics experiments suggest that ant kooperative effects in the presence of guanosine 5′-monophosphate arise primarily from slower formation of tertiary contacts between the catalytic core of the ribozyme and the P1 duplex formed by pyCUCU and GGAGGG of the ribozyme. Conversely, cooperative effects in the presence of 2′-deoxyguanosine 5′-monophosphate arise primarily from slower disruption of tertiary contacts between the catalytic core of the ribozyme and the P1 duplex. Additional experiments suggest that these cooperative and anticooperative effects are not a function of the pyrene label, are not caused by a salt effect, and are not specific to one renaturation procedure for the ribozyme.",
author = "Bevilacqua, {Philip C.} and Johnson, {Kenneth A.} and Turner, {Douglas H.}",
year = "1993",
month = "9",
day = "15",
doi = "10.1073/pnas.90.18.8357",
language = "English (US)",
volume = "90",
pages = "8357--8361",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "18",

}

Cooperative and anticooperative binding to a ribozyme. / Bevilacqua, Philip C.; Johnson, Kenneth A.; Turner, Douglas H.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 18, 15.09.1993, p. 8357-8361.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cooperative and anticooperative binding to a ribozyme

AU - Bevilacqua, Philip C.

AU - Johnson, Kenneth A.

AU - Turner, Douglas H.

PY - 1993/9/15

Y1 - 1993/9/15

N2 - The effects of guanosine 5′-monophosphate and 2′-deoxyguanosine 5′-monophosphate on the thermodynamics and kinetics of pyrene-labeled 5′ exon mimic (pyCUCU) binding to the catalytic RNA (ribozyme) from Tetrahymena thermophila have been determined by fluorescence titration and kinetics experiments at 15°C. pyCUCU binding to L-21 Sca I-truncated ribozyme is weaker by a factor of 5 in the presence of saturating guanosine 5′-monophosphate, whereas it is 4-fold stronger in the presence of saturating 2′-deoxyguanosine 5′-monophosphate. Results from kinetics experiments suggest that ant kooperative effects in the presence of guanosine 5′-monophosphate arise primarily from slower formation of tertiary contacts between the catalytic core of the ribozyme and the P1 duplex formed by pyCUCU and GGAGGG of the ribozyme. Conversely, cooperative effects in the presence of 2′-deoxyguanosine 5′-monophosphate arise primarily from slower disruption of tertiary contacts between the catalytic core of the ribozyme and the P1 duplex. Additional experiments suggest that these cooperative and anticooperative effects are not a function of the pyrene label, are not caused by a salt effect, and are not specific to one renaturation procedure for the ribozyme.

AB - The effects of guanosine 5′-monophosphate and 2′-deoxyguanosine 5′-monophosphate on the thermodynamics and kinetics of pyrene-labeled 5′ exon mimic (pyCUCU) binding to the catalytic RNA (ribozyme) from Tetrahymena thermophila have been determined by fluorescence titration and kinetics experiments at 15°C. pyCUCU binding to L-21 Sca I-truncated ribozyme is weaker by a factor of 5 in the presence of saturating guanosine 5′-monophosphate, whereas it is 4-fold stronger in the presence of saturating 2′-deoxyguanosine 5′-monophosphate. Results from kinetics experiments suggest that ant kooperative effects in the presence of guanosine 5′-monophosphate arise primarily from slower formation of tertiary contacts between the catalytic core of the ribozyme and the P1 duplex formed by pyCUCU and GGAGGG of the ribozyme. Conversely, cooperative effects in the presence of 2′-deoxyguanosine 5′-monophosphate arise primarily from slower disruption of tertiary contacts between the catalytic core of the ribozyme and the P1 duplex. Additional experiments suggest that these cooperative and anticooperative effects are not a function of the pyrene label, are not caused by a salt effect, and are not specific to one renaturation procedure for the ribozyme.

UR - http://www.scopus.com/inward/record.url?scp=0027239573&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027239573&partnerID=8YFLogxK

U2 - 10.1073/pnas.90.18.8357

DO - 10.1073/pnas.90.18.8357

M3 - Article

VL - 90

SP - 8357

EP - 8361

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 18

ER -