Rz/Rz1 Lysis Gene Equivalents in Phages of Gram-negative Hosts

Summer, Elizabeth J.; Berry, Joel D.; Tran, Tri M.; Niu, Lili; Struck, Douglas K.; Young, Ry

doi:10.1016/j.jmb.2007.08.045

Cited by 148 publications

(210 citation statements)

References 48 publications

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“…The predicted N-terminal TMD and a-helical regions are highlighted and underlined, respectively. 8 The region of Rz deleted during the construction of pSRzH 6 (see ''Materials and methods'') is indicated by a cross bar and D symbol. The sequence and location of the C-terminal oligohistidine tag are also indicated.…”

Section: Purification Of the Rz And Rz1 Periplasmic Domainsmentioning

confidence: 99%

“…Both topology and genetics mandate that this interaction occurs via the C-termini of both proteins. 1,8 Evidence that this interaction occurs in vivo was obtained by assessing the state of the full-length Rz protein in whole cells in which the Rz gene was expressed at physiological levels ( Fig. 5).…”

Section: The Rz Protein Is Stabilized By Rz1 In Vivomentioning

confidence: 99%

“…6,7 Aided by the striking architecture, which is unique in biology for two genes involved in the same function, we identified equivalents of Rz-Rz1 in nearly all genomes of phages that infect Gram-negative bacteria. 8 The ubiquity and diversity (37 unrelated gene families) suggested that, for phage lysis, Rz and Rz1 are just as important as holins and endolysins.…”

Section: Introductionmentioning

confidence: 99%

“…1(A)]. 8 The mature Rz1 protein is a 40 residue OM lipoprotein containing 10 Pro residues and devoid of predicted secondary structure [ Fig. 1(B)].…”

Section: Introductionmentioning

confidence: 99%

“…1(B)]. 8,10 Multiple lines of genetic, physiological, and molecular evidence suggest that Rz and Rz1 form a complex, which, as it must span the periplasm, has been designated as the spanin complex. According to this formalism, Rz and Rz1 are the large and small spanin subunits, respectively.…”

Section: Introductionmentioning

confidence: 99%

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The lambda spanin components Rz and Rz1 undergo tertiary and quaternary rearrangements upon complex formation

et al. 2010

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Phage holins and endolysins have long been known to play key roles in lysis of the host cell, disrupting the cytoplasmic membrane and peptidoglycan (PG) layer, respectively. For phages of Gram-negative hosts, a third class of proteins, the spanins, are involved in disrupting the outer membrane (OM). Rz and Rz1, the components of the lambda spanin, are, respectively, a class II inner membrane protein and an OM lipoprotein, are thought to span the entire periplasm by virtue of C-terminal interactions of their soluble domains. Here, the periplasmic domains of Rz and Rz1 have been purified and shown to form dimeric and monomeric species, respectively, in solution. Circular dichroism analysis indicates that Rz has significant alpha-helical character, but much less than predicted, whereas Rz1, which is 25% proline, is unstructured. Mixture of the two proteins leads to complex formation and an increase in secondary structure, especially alpha-helical content. Moreover, transmission electron-microscopy reveals that Rz-Rz1 complexes form large rod-shaped structures which, although heterogeneous, exhibit periodicities that may reflect coiledcoil bundling as well as a long dimension that matches the width of the periplasm. A model is proposed suggesting that the formation of such bundles depends on the removal of the PG and underlies the Rz-Rz1 dependent disruption of the OM.

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Section: Purification Of the Rz And Rz1 Periplasmic Domainsmentioning

confidence: 99%

Section: The Rz Protein Is Stabilized By Rz1 In Vivomentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…1(A)]. 8 The mature Rz1 protein is a 40 residue OM lipoprotein containing 10 Pro residues and devoid of predicted secondary structure [ Fig. 1(B)].…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The lambda spanin components Rz and Rz1 undergo tertiary and quaternary rearrangements upon complex formation

et al. 2010

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Phage Genome Annotation Using the RAST Pipeline

McNair

Aziz

Pusch

et al. 2017

Methods in Molecular Biology

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Phages are complex biomolecular machineries that have to survive in a bacterial world. Phage genomes show many adaptations to their lifestyle such as shorter genes, reduced capacity for redundant DNA sequences, and the inclusion of tRNAs in their genomes. In addition, phages are not free-living, they require a host for replication and survival. These unique adaptations provide challenges for the bioinformatics analysis of phage genomes. In particular, ORF calling, genome annotation, noncoding RNA (ncRNA) identification, and the identification of transposons and insertions are all complicated in phage genome analysis. We provide a road map through the phage genome annotation pipeline, and discuss the challenges and solutions for phage genome annotation as we have implemented in the rapid annotation using subsystems (RAST) pipeline.

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Analysis of the cell wall binding domain in bacteriocin-like lysin LysL from Lactococcus lactis LAC460

Mokhtari,

Li,

Saris

et al. 2024

Arch Microbiol

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Wild-type Lactococcus lactis strain LAC460 secretes prophage-encoded bacteriocin-like lysin LysL, which kills some Lactococcus strains, but has no lytic effect on the producer. LysL carries two N-terminal enzymatic active domains (EAD), and an unknown C-terminus without homology to known domains. This study aimed to determine whether the C-terminus of LysL carries a cell wall binding domain (CBD) for target specificity of LysL. The C-terminal putative CBD region of LysL was fused with His-tagged green fluorescent protein (HGFPuv). The HGFPuv_CBDlysL gene fusion was ligated into the pASG-IBA4 vector, and introduced into Escherichia coli. The fusion protein was produced and purified with affinity chromatography. To analyse the binding of HGFPuv_CBDLysL to Lactococcus cells, the protein was mixed with LysL-sensitive and LysL-resistant strains, including the LysL-producer LAC460, and the fluorescence of the cells was analysed. As seen in fluorescence microscope, HGFPuv_CBDLysL decorated the cell surface of LysL-sensitive L. cremoris MG1614 with green fluorescence, whereas the resistant L. lactis strains LM0230 and LAC460 remained unfluorescent. The fluorescence plate reader confirmed the microscopy results detecting fluorescence only from four tested LysL-sensitive strains but not from 11 tested LysL-resistant strains. Specific binding of HGFPuv_CBDLysL onto the LysL-sensitive cells but not onto the LysL-resistant strains indicates that the C-terminus of LysL contains specific CBD. In conclusion, this report presents experimental evidence of the presence of a CBD in a lactococcal phage lysin. Moreover, the inability of HGFPuv_CBDLysL to bind to the LysL producer LAC460 may partly explain the host’s resistance to its own prophage lysin.

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Rz/Rz1 Lysis Gene Equivalents in Phages of Gram-negative Hosts

Cited by 148 publications

References 48 publications

The lambda spanin components Rz and Rz1 undergo tertiary and quaternary rearrangements upon complex formation

The lambda spanin components Rz and Rz1 undergo tertiary and quaternary rearrangements upon complex formation

Phage Genome Annotation Using the RAST Pipeline

Analysis of the cell wall binding domain in bacteriocin-like lysin LysL from Lactococcus lactis LAC460

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