A possible role for RNA polymerase in the initiation of M13 DNA synthesis.

نویسندگان

  • D Brutlag
  • R Schekman
  • A Kornberg
چکیده

The conversion of single-stranded DNA of bacteriophage M13 to the double-stranded replicative form in Escherichia coli is blocked by rifampicin, an antibiotic that specifically inhibits the host-cell RNA polymerase. Chloramphenicol, an inhibitor of protein synthesis, does not block this conversion. The next stage in phage DNA replication, multiplication of the doublestranded forms, is also inhibited by rifampicin; chloramphenicol, although inhibitory, has a much smaller effect. An E. coli mutant whose RNA polymerase is resistant to rifampicin action does not show inhibition of M13 DNA replication by rifampicin. These findings indicate that a specific rifampicin-RNA polymerase interaction is responsible for blocking new DNA synthesis. It now seems plausible that RNA polymerase has some direct role in the initiation of DNA replication, perhaps by forming a primer RNA that serves for covalent attachment of the deoxyribonucleotide that starts the new DNA chain.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

The role of RNA polymerase subunit in promoter-independent initiation of transcription

In bacteria, initiation of transcription depends on the RNA polymerase subunit, which brings catalytically proficient RNA polymerase core to promoters by binding to specific DNA elements located upstream of the transcription start point. Here, we study -dependent synthesis of a transcript that is used to prime replication of the single-stranded genome of bacteriophage M13. We show that, in this...

متن کامل

Further biochemical characterization of wheat DNA primase: possible functional implication of copurification with DNA polymerase A.

DNA primase has been partially purified from wheat germ. This enzyme, like DNA primases characterized from many procaryotic and eucaryotic sources, catalyses the synthesis of primers involved in DNA replication. However, the wheat enzyme differs from animal DNA primase in that it is found partially associated with a DNA polymerase which differs greatly from DNA polymerase alpha. Moreover, the o...

متن کامل

RNA Synthesis Initiates In Vitro Conversion of M 1 3 DNA to Its Replicative Form ( 6 x 174 DNA / DNA initiation / DNA replication / rifampicin )

Soluble enzyme fractions from uninfected Escherichia coli convert M13 and 6x174 viral single strands to their doublestranded replicative forms. Rifampicin, an inhibitor of RNA polymerase, blocks conversion of M13 single strands to the replicative forms in vivo and in vitro. However, rifampicin does not block synthesis of the replicative forms of 6x174 either in vivo or in soluble extracts. The ...

متن کامل

Selective cloning of a DNA single-strand initiation determinant from phi X174 replicative-form DNA.

An M13 phage deletion mutant, M13 delta E101, developed as a vector for selecting DNA sequences that direct DNA strand initiation on a single-stranded template, has been used for cloning restriction enzyme digests of phi X174 replicative-form DNA. Initiation determinants, detected on the basis of clear-plaque formation by the chimeric phage, were found only in restriction fragments containing t...

متن کامل

A Novel Form of RNA Polymerase from Escherichia coli ( M 13 / + X 174 / rifampicin )

A new form of RNA polymerase, termed RNA polymerase 111, has been recognized as a large fraction of the rifampicin-sensitive enzyme in E. coli. It is physically separable from RNA polymerase (holoenzyme, RNA polymerase I) by gel filtration and is distinguished by its capacity to discriminate between M13 and 4x174 viral DNA templates in priming DNA synthesis. This template specificity is manifes...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 68 11  شماره 

صفحات  -

تاریخ انتشار 1971