The Ordered Assembly of the φX174-type Primosome

Liu, Joing; Nurse, Pearl; Marians, Kenneth J.

doi:10.1074/jbc.271.26.15656

Cited by 77 publications

(53 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, we postulate that PriB binds ssDNA that has been unwound or presented by PriA and then positions or maintains the bound ssDNA in a specific conformation that allows DnaT recognition and binding. This hypothesis is consistent with the data showing that in the presence of excess DnaT, PriB is not required for the in vitro primosome assembly (19).…”

Section: And B)supporting

confidence: 93%

“…The SSB-like fold may help PriB in displacing SSB from ssDNA (25). It is known that DnaT interacts with the PriA-PriB-DNA complex during primosome assembly (16,19) and that DnaT is capable of binding ssDNA (48). Therefore, we postulate that PriB binds ssDNA that has been unwound or presented by PriA and then positions or maintains the bound ssDNA in a specific conformation that allows DnaT recognition and binding.…”

Section: And B)mentioning

confidence: 99%

“…One could speculate that the region containing Cys-12 or Cys-27 may be a potential protein-protein interface for PriB to interact with other proteins during primosome assembly. According to the current knowledge of primosome assembly, PriA and DnaT are both candidates that can physically interact with PriB (16,19). DnaT has no cysteine in its polypeptide sequence (48) and, therefore, it is unlikely to interact with PriB through intermolecular disulfide bridging.…”

Section: And B)mentioning

confidence: 99%

“…PriA has two cysteine residues, Cys-439 and Cys-445. These cysteine residues are required for the formation of the PriA-PriB complex (19,54). However, it is still unclear how these cysteine residues participate in the PriA-PriB complex formation.…”

Section: And B)mentioning

confidence: 99%

“…During the formation of the preprimosome assembly, PriB interacts with and stabilizes the PriA-DNA complex (10,16), facilitating the complex formation between PriA and DnaT (19). PriB was formerly known as the "n protein" because it can be inactivated by treatment with N-ethylmaleimide (NEM) (13).…”

mentioning

confidence: 99%

See 4 more Smart Citations

Crystal Structure of PriB, a Primosomal DNA Replication Protein of Escherichia coli

Liu

Chang²,

Huang

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

PriB is one of the Escherichia coli X-type primosome proteins that are required for assembly of the primosome, a mobile multi-enzyme complex responsible for the initiation of DNA replication. Here we report the crystal structure of the E. coli PriB at 2.1 Å resolution by multi-wavelength anomalous diffraction using a mercury derivative. The polypeptide chain of PriB is structurally similar to that of single-stranded DNA-binding protein (SSB). However, the biological unit of PriB is a dimer, not a homotetramer like SSB. Electrophoretic mobility shift assays demonstrated that PriB binds single-stranded DNA and single-stranded RNA with comparable affinity. We also show that PriB binds singlestranded DNA with certain base preferences. Based on the PriB structural information and biochemical studies, we propose that the potential tetramer formation surface and several other regions of PriB may participate in protein-protein interaction during DNA replication. These findings may illuminate the role of PriB in X-type primosome assembly.

show abstract

Section: And B)supporting

confidence: 93%

Section: And B)mentioning

confidence: 99%

Section: And B)mentioning

confidence: 99%

Section: And B)mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Crystal Structure of PriB, a Primosomal DNA Replication Protein of Escherichia coli

Liu

Chang²,

Huang

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

A structural model of the PriB–DnaT complex in Escherichia coli replication restart

et al. 2020

View full text Add to dashboard Cite

In Escherichia coli, DNA replication is restarted following DNA repair by the PriA‐dependent pathway, in which the binding and dissociation of proteins such as PriA, PriB, and DnaT on ssDNA lead to the formation of a protein–DNA complex for recruiting the DnaB–DnaC replication protein complex. However, the structure of the PriB–DnaT complex, which is an essential step in the PriA‐dependent pathway, remains elusive. In this study, the importance of His26 in PriB for replication restart was reconfirmed using plasmid complementation. Furthermore, we used NMR to examine the DnaT interaction sites on PriB. We also evaluated the PriB–DnaT peptide complex model, which was prepared by in silico docking, using molecular dynamic simulation. From these data, we propose a structural model that provides insight into the PriB–DnaT interaction.

show abstract

Bacterial Primosome

Tanaka¹,

Masai²

2019

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

DNA (deoxyribonucleic acid) replication requires operation of molecular machinery which efficiently elongates nucleotide chains on both strands. This process requires not only the enzymes synthesising DNA (DNA polymerases) but also those providing primer RNAs (ribonucleic acid) and continuously melting the duplex DNA. The primosome refers to a protein complex capable of processive unwinding of duplex DNA and primer RNA synthesis on the lagging strand at a replication fork. The prepriming proteins, DNA helicase and primase are sequentially assembled on the template DNA to generate a primosome. Once assembled, it, in conjunction with DNA polymerases, facilitates DNA chain elongation. The assembly of bacterial primosome is triggered by an ‘initiator’ protein including DnaA or PriA, which recognises the site of assembly. Primosome is assembled also in replication restart process at stalled or processed replication forks, triggered by PriA. Primosome constitutes an essential component for active replication fork machinery. Recent advances in this field provide structural basis regarding how these factors function during the assembly of a primosome on viral origins as well as during the restart from the stalled DNA replication forks. These new knowledges provide important insight into how replication forks are protected from various genotoxic agents in eukaryotes. Key Concepts Replication fork is the site of DNA replication where DNA synthesis occurs, and primosome is its integral component. Primosome is capable of duplex DNA unwinding and primer RNA synthesis at the replication fork. In bacteria, replication is normally initiated at a single locus, oriC , on the genome. Bacterial DNA replication is initiated by an initiator, DnaA, which generates a oriC ‐primosome. The stalled DNA replication fork needs to be swiftly detected and rescued to prevent its collapse and to ensure the completion of genome replication. Another primosome mediated by PriA serves for reassembly of replication fork at a stalled fork in bacteria. Bacterial genomes can be replicated by an alternative mode that involves RNA‐DNA hybrids. Primosomes in eukaryotes may be more complex, but essential components would be conserved.

show abstract

The Ordered Assembly of the φX174-type Primosome

Cited by 77 publications

References 23 publications

Crystal Structure of PriB, a Primosomal DNA Replication Protein of Escherichia coli

Crystal Structure of PriB, a Primosomal DNA Replication Protein of Escherichia coli

A structural model of the PriB–DnaT complex in Escherichia coli replication restart

Bacterial Primosome

Contact Info

Product

Resources

About