Archives of Microbiology

, Volume 181, Issue 4, pp 305–313 | Cite as

Lagging strand replication of rolling-circle plasmids in Streptomyces lividans: an RNA polymerase-independent primer synthesis

  • Ichiro Suzuki
  • Masakazu KataokaEmail author
  • Toshiomi Yoshida
  • Tatsuji Seki
Original Paper


The rolling circle (RC) mechanism of DNA replication generating single-stranded DNA (ssDNA) intermediates is common in various high-copy circular plasmids in Streptomyces, and the ssDNA released after leading strand synthesis is converted to its double-stranded form (dsDNA) by the host proteins. The in vivo and in vitro lagging strand syntheses from ssDNA replicative intermediates of RC plasmid pSN22 in Streptomyces lividans was characterized. The presence or absence of the single-strand origin (sso), the replication initiation site of lagging strand synthesis, did not significantly affect the copy numbers of pSN22 derivatives. In vivo lagging strand synthesis was not affected by the rifampicin inhibition of S. lividans RNA polymerase. Likewise, in vitro lagging strand synthesis using cell-free extracts revealedsso-independent, rifampicin-resistant lagging strand synthesis in S. lividans. Although all four dNTPs are usually required for the initiation of such synthesis, the presence of only one NTP was sufficient to carry outlagging strand synthesis in vitro. Interestingly, the cell-free extract of exponential-phase cells required less ATP than that of stationary-phase cells. These results reveal a predominant RNA polymerase-independent priming system in S. lividans that may be a result of the stabilization of RC plasmids lacking sso in S. lividans.


Replication Streptomyces Priming Lagging strand 



We greatly thank to Drs. T. Kieser, H. Araki and T. Ito for valuable discussions; Dr E. Ko-Mitamura for editorial suggestions and manuscript corrections; and Dr H. Masai for valuable comments on the general priming of E. coli.


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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Ichiro Suzuki
    • 1
    • 2
  • Masakazu Kataoka
    • 1
    • 3
    Email author
  • Toshiomi Yoshida
    • 1
  • Tatsuji Seki
    • 1
  1. 1.The International Center for BiotechnologyOsaka UniversityOsakaJapan
  2. 2.Department of Chemical Engineering Science, Graduate School of EngineeringYokohama National UniversityYokohamaJapan
  3. 3.Department of Environmental Science and Technology, Faculty of EngineeringShinshu UniversityNaganoJapan

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