A 39 amino acid fragment of the cell cycle regulator p21 is sufficient to bind PCNA and partially inhibit DNA replication in vivo

Junjie Chen; Richard Peters; Partha Saha; Patrick Lee; Annie Theodoras; Michele Pagano; Gerhard Wagner; Anindya Dutta

doi:10.1093/nar/24.9.1727

A 39 amino acid fragment of the cell cycle regulator p21 is sufficient to bind PCNA and partially inhibit DNA replication in vivo

Junjie Chen, Richard Peters, Partha Saha, Patrick Lee, Annie Theodoras, Michele Pagano, Gerhard Wagner, Anindya Dutta

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Resumen

The cell cycle regulator p21 interacts with and inhibits the DNA replication and repair factor proliferating cell nuclear antigen (PCNA). We have defined a 39 amino acid fragment of p21 which is sufficient to bind PCNA with high affinity (K(d) 10-20 nM). This peptide can inhibit DNA replication in vitro and microinjection of a GST fusion protein containing this domain inhibited S phase in vivo. Despite its high affinity for PCNA, the free 39 amino acid peptide does not have a well-defined structure, as judged from circular dichroism and nuclear magnetic resonance measurements, suggesting an induced fit mechanism for the PCNA-p21 interaction. The association of the small peptide with PCNA was thermolabile, suggesting that portions of p21 adjoining the minimal region of contact stabilize the interaction. In addition, a domain containing 67 amino acids from the N-terminus of PCNA was defined as both necessary and sufficient for binding to p21.

Idioma original	English
Páginas (desde-hasta)	1727-1733
Número de páginas	7
Publicación	Nucleic Acids Research
Volumen	24
N.º	9
DOI	https://doi.org/10.1093/nar/24.9.1727
Estado	Published - 1996
Publicado de forma externa	Sí

Nota bibliográfica

Funding Information:
We thank members of the Dutta Laboratory and G.Lindenmeyer and D.Gilbert for helpful discussions, J.Morrow for technical assistance, J.Parvin for reading the manuscript, C.Dahl for help with peptide synthesis and J.Lee for use of the CD spectrometer. This work was supported by a grant from the NIH (CA60499) and career development awards from the American Cancer Society (JFRA 474) and the US Armed Forces Medical Research Command (DAMD17-94-J-4064). JC was supported by a post-doctoral fellowship (DAMD 17-94-J-4070), RP is a Howard Hughes Medical Institute Physician Post-doctoral Fellow, and MP was supported in part by HSFP grant RG-496/93.

ASJC Scopus Subject Areas

Genetics

PubMed: MeSH publication types

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

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10.1093/nar/24.9.1727

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title = "A 39 amino acid fragment of the cell cycle regulator p21 is sufficient to bind PCNA and partially inhibit DNA replication in vivo",

abstract = "The cell cycle regulator p21 interacts with and inhibits the DNA replication and repair factor proliferating cell nuclear antigen (PCNA). We have defined a 39 amino acid fragment of p21 which is sufficient to bind PCNA with high affinity (K(d) 10-20 nM). This peptide can inhibit DNA replication in vitro and microinjection of a GST fusion protein containing this domain inhibited S phase in vivo. Despite its high affinity for PCNA, the free 39 amino acid peptide does not have a well-defined structure, as judged from circular dichroism and nuclear magnetic resonance measurements, suggesting an induced fit mechanism for the PCNA-p21 interaction. The association of the small peptide with PCNA was thermolabile, suggesting that portions of p21 adjoining the minimal region of contact stabilize the interaction. In addition, a domain containing 67 amino acids from the N-terminus of PCNA was defined as both necessary and sufficient for binding to p21.",

author = "Junjie Chen and Richard Peters and Partha Saha and Patrick Lee and Annie Theodoras and Michele Pagano and Gerhard Wagner and Anindya Dutta",

note = "Funding Information: We thank members of the Dutta Laboratory and G.Lindenmeyer and D.Gilbert for helpful discussions, J.Morrow for technical assistance, J.Parvin for reading the manuscript, C.Dahl for help with peptide synthesis and J.Lee for use of the CD spectrometer. This work was supported by a grant from the NIH (CA60499) and career development awards from the American Cancer Society (JFRA 474) and the US Armed Forces Medical Research Command (DAMD17-94-J-4064). JC was supported by a post-doctoral fellowship (DAMD 17-94-J-4070), RP is a Howard Hughes Medical Institute Physician Post-doctoral Fellow, and MP was supported in part by HSFP grant RG-496/93.",

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doi = "10.1093/nar/24.9.1727",

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journal = "Nucleic Acids Research",

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AU - Chen, Junjie

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AU - Saha, Partha

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AU - Theodoras, Annie

AU - Pagano, Michele

AU - Wagner, Gerhard

AU - Dutta, Anindya

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PY - 1996

Y1 - 1996

N2 - The cell cycle regulator p21 interacts with and inhibits the DNA replication and repair factor proliferating cell nuclear antigen (PCNA). We have defined a 39 amino acid fragment of p21 which is sufficient to bind PCNA with high affinity (K(d) 10-20 nM). This peptide can inhibit DNA replication in vitro and microinjection of a GST fusion protein containing this domain inhibited S phase in vivo. Despite its high affinity for PCNA, the free 39 amino acid peptide does not have a well-defined structure, as judged from circular dichroism and nuclear magnetic resonance measurements, suggesting an induced fit mechanism for the PCNA-p21 interaction. The association of the small peptide with PCNA was thermolabile, suggesting that portions of p21 adjoining the minimal region of contact stabilize the interaction. In addition, a domain containing 67 amino acids from the N-terminus of PCNA was defined as both necessary and sufficient for binding to p21.

AB - The cell cycle regulator p21 interacts with and inhibits the DNA replication and repair factor proliferating cell nuclear antigen (PCNA). We have defined a 39 amino acid fragment of p21 which is sufficient to bind PCNA with high affinity (K(d) 10-20 nM). This peptide can inhibit DNA replication in vitro and microinjection of a GST fusion protein containing this domain inhibited S phase in vivo. Despite its high affinity for PCNA, the free 39 amino acid peptide does not have a well-defined structure, as judged from circular dichroism and nuclear magnetic resonance measurements, suggesting an induced fit mechanism for the PCNA-p21 interaction. The association of the small peptide with PCNA was thermolabile, suggesting that portions of p21 adjoining the minimal region of contact stabilize the interaction. In addition, a domain containing 67 amino acids from the N-terminus of PCNA was defined as both necessary and sufficient for binding to p21.

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A 39 amino acid fragment of the cell cycle regulator p21 is sufficient to bind PCNA and partially inhibit DNA replication in vivo

Resumen

Nota bibliográfica

ASJC Scopus Subject Areas

PubMed: MeSH publication types

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