Deoxyribonucleic acid chain growth and organization of replicating units in HeLa cells

Stephen Planck, Gerald C. Mueller

Research output: Contribution to journalArticle

14 Citations (Scopus)

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 replicons 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 immediate 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 languageEnglish (US)
Pages (from-to)1808-1813
Number of pages6
JournalBiochemistry
Volume16
Issue number9
StatePublished - 1977
Externally publishedYes

Fingerprint

HeLa Cells
Replicon
DNA
Bromodeoxyuridine
Growth
DNA Replication
Thymidine
Photolysis
Cycloheximide
S Phase
Sucrose
Sedimentation
Labeling
Light
Temperature
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Deoxyribonucleic acid chain growth and organization of replicating units in HeLa cells. / Planck, Stephen; Mueller, Gerald C.

In: Biochemistry, Vol. 16, No. 9, 1977, p. 1808-1813.

Research output: Contribution to journalArticle

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