Deoxyribonucleic Acid Chain Growth and Organization of Replicating Units in HeLa Cells

Stephen R. Planck; Gerald C. Mueller

doi:10.1021/bi00628a007

Deoxyribonucleic Acid Chain Growth and Organization of Replicating Units in HeLa Cells

Stephen R. Planck, Gerald C. Mueller

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

A method for studying DNA chain growth and chromosomal organization of replicons in HeLa cells has been developed. DNA replication is initiated with bromodeoxyuridine followed by pulse labeling of active replicorts With [³H]thymidine and growth of the chains for finite intervals in unlabeled thymidine. Photolysis of the bromodeoxyuridine-DNA leader with 313-nm light releases the newly replicated chains that are then analyzed by sedimentation in alkaline sucrose gradients. This method of analysis provides data on the rate of chain growth, the bidirectionality of replication, and the distribution of the active replicons at specific intervals in the S period. Applying this method to cells caused to synthesize DNA at a lowered temperature (27 °C) or with protein synthesis restricted by cycloheximide revealed that the irrimediate reduction in the rate of DNA replication in both instances was due to a decreased rate of chain growth without derangement of the overall process.

Original language	English (US)
Pages (from-to)	1808-1813
Number of pages	6
Journal	Biochemistry
Volume	16
Issue number	9
DOIs	https://doi.org/10.1021/bi00628a007
State	Published - May 1 1977
Externally published	Yes

ASJC Scopus subject areas

Biochemistry

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10.1021/bi00628a007

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title = "Deoxyribonucleic Acid Chain Growth and Organization of Replicating Units in HeLa Cells",

abstract = "A method for studying DNA chain growth and chromosomal organization of replicons in HeLa cells has been developed. DNA replication is initiated with bromodeoxyuridine followed by pulse labeling of active replicorts With [3H]thymidine and growth of the chains for finite intervals in unlabeled thymidine. Photolysis of the bromodeoxyuridine-DNA leader with 313-nm light releases the newly replicated chains that are then analyzed by sedimentation in alkaline sucrose gradients. This method of analysis provides data on the rate of chain growth, the bidirectionality of replication, and the distribution of the active replicons at specific intervals in the S period. Applying this method to cells caused to synthesize DNA at a lowered temperature (27 °C) or with protein synthesis restricted by cycloheximide revealed that the irrimediate reduction in the rate of DNA replication in both instances was due to a decreased rate of chain growth without derangement of the overall process.",

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year = "1977",

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volume = "16",

pages = "1808--1813",

journal = "Biochemistry",

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publisher = "American Chemical Society",

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TY - JOUR

T1 - Deoxyribonucleic Acid Chain Growth and Organization of Replicating Units in HeLa Cells

AU - Planck, Stephen R.

AU - Mueller, Gerald C.

PY - 1977/5/1

Y1 - 1977/5/1

N2 - A method for studying DNA chain growth and chromosomal organization of replicons in HeLa cells has been developed. DNA replication is initiated with bromodeoxyuridine followed by pulse labeling of active replicorts With [3H]thymidine and growth of the chains for finite intervals in unlabeled thymidine. Photolysis of the bromodeoxyuridine-DNA leader with 313-nm light releases the newly replicated chains that are then analyzed by sedimentation in alkaline sucrose gradients. This method of analysis provides data on the rate of chain growth, the bidirectionality of replication, and the distribution of the active replicons at specific intervals in the S period. Applying this method to cells caused to synthesize DNA at a lowered temperature (27 °C) or with protein synthesis restricted by cycloheximide revealed that the irrimediate reduction in the rate of DNA replication in both instances was due to a decreased rate of chain growth without derangement of the overall process.

AB - A method for studying DNA chain growth and chromosomal organization of replicons in HeLa cells has been developed. DNA replication is initiated with bromodeoxyuridine followed by pulse labeling of active replicorts With [3H]thymidine and growth of the chains for finite intervals in unlabeled thymidine. Photolysis of the bromodeoxyuridine-DNA leader with 313-nm light releases the newly replicated chains that are then analyzed by sedimentation in alkaline sucrose gradients. This method of analysis provides data on the rate of chain growth, the bidirectionality of replication, and the distribution of the active replicons at specific intervals in the S period. Applying this method to cells caused to synthesize DNA at a lowered temperature (27 °C) or with protein synthesis restricted by cycloheximide revealed that the irrimediate reduction in the rate of DNA replication in both instances was due to a decreased rate of chain growth without derangement of the overall process.

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JO - Biochemistry

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ER -

Deoxyribonucleic Acid Chain Growth and Organization of Replicating Units in HeLa Cells

Abstract

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