Theta replication of the lactococcal plasmid pWVO2

Kiewiet, Rense; Bron, Sierd; Jonge, Karel de; Venema, Gerard; Seegers, Jos F. M. L.

doi:10.1111/j.1365-2958.1993.tb01958.x

Cited by 84 publications

(87 citation statements)

References 40 publications

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“…The complementation of prsA3 was determined with IH7492 (prsA3 trpC2 glyB133 hisA1), which expresses amyQ of Bacillus amyloliquefaciens from pKTH3382. The pKTH3382 plasmid was constructed by inserting the amyQ gene as a SalI fragment into pSR11 (22). IH7492 was also used as the parental strain to produce and purify various lipomodified PrsA proteins.…”

Section: Methodsmentioning

confidence: 99%

Structure-Function Analysis of PrsA Reveals Roles for the Parvulin-like and Flanking N- and C-terminal Domains in Protein Folding and Secretion in Bacillus subtilis

Vitikainen

Lappalainen

Seppälä

et al. 2004

Journal of Biological Chemistry

118

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The PrsA protein of Bacillus subtilis is an essential membrane-bound lipoprotein that is assumed to assist post-translocational folding of exported proteins and stabilize them in the compartment between the cytoplasmic membrane and cell wall. This folding activity is consistent with the homology of a segment of PrsA with parvulin-type peptidyl-prolyl cis/trans isomerases (PPIase). In this study, molecular modeling showed that the parvulin-like region can adopt a parvulin-type fold with structurally conserved active site residues. PrsA exhibits PPIase activity in a manner dependent on the parvulin-like domain. We constructed deletion, peptide insertion, and amino acid substitution mutations and demonstrated that the parvulin-like domain as well as flanking N-and C-terminal domains are essential for in vivo PrsA function in protein secretion and growth. Surprisingly, none of the predicted active site residues of the parvulin-like domain was essential for growth and protein secretion, although several active site mutations reduced or abolished the PPIase activity or the ability of PrsA to catalyze proline-limited protein folding in vitro. Our results indicate that PrsA is a PPIase, but the essential role in vivo seems to depend on some non-PPIase activity of both the parvulin-like and flanking domains.

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Section: Methodsmentioning

confidence: 99%

Structure-Function Analysis of PrsA Reveals Roles for the Parvulin-like and Flanking N- and C-terminal Domains in Protein Folding and Secretion in Bacillus subtilis

Vitikainen

Lappalainen

Seppälä

et al. 2004

Journal of Biological Chemistry

118

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“…To date, the replication regions of more than 20 lactococcal plasmids have been sequenced and have been shown to belong to the highly homologous pCI305-type family (21), of which pWV02 has been shown to replicate via the mode (27). On the basis of homologies and structural organization, these plasmids can be considered class A plasmids.…”

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confidence: 99%

Identification and Characterization of an Active Plasmid Partition Mechanism for the Novel Lactococcus lactis Plasmid pCI2000

2000

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The replication region of the lactococcal plasmid pCI2000 was subcloned and analyzed. The nucleotide sequence of one 5.6-kb EcoRI fragment which was capable of supporting replication when cloned on a replication probe vector revealed the presence of seven putative open reading frames (ORFs). One ORF exhibited significant homology to several replication proteins from plasmids considered to replicate via a theta mode. Deletion analysis showed that this ORF, designated repA, is indeed required for replication. The results also suggest that the origin of replication is located outside repA. Upstream and divergently transcribed from repA, an ORF that showed significant (48 to 64%) homology to a number of proteins that are required for faithful segregation of chromosomal or plasmid DNA of gram-negative bacteria was identified. Gene interruption and transcomplementation experiments showed that this ORF, designated parA, is required for stable inheritance of pCI2000 and is active in trans. This is the first example of such a partitioning mechanism for plasmids in gram-positive bacteria.Lactococcus lactis strains are of considerable industrial and economic importance, as they are widely used in the production of a variety of fermented dairy products. A characteristic of Lactococcus strains is that they typically possess an abundance of plasmid DNA on which a number of significant technological traits are encoded. These include bacteriophage resistance, bacteriocin production, lactose assimilation, citrate utilization, and proteinase activity (11). Therefore, an extensive knowledge of lactococcal plasmid replication, partition, and stability functions is essential in order to ensure the stable maintenance of these traits. This information can also be applied for the generation of novel stable food-grade cloning and expression vectors for the manipulation of these hosts.There are two modes of bacterial plasmid replication, rolling circle (RC) and theta ( ), both of which have been identified in Lactococcus. The RC plasmids are classified on the basis of homologies within the region of the double-stranded origin and the gene encoding the replication initiation protein. Two classes of RC plasmid are evident in Lactococcus, pE194-like and pC194-like, of which pWV01 (26, 28) and pWC1 (33), respectively, are the prototypes. In general, replicons are classified according to their structural organization and the requirement for host-encoded proteins in the replication process. Originally, the best-characterized replicating plasmids were of gram-negative origin, and three classes designated A, B, and C were identified. Class A plasmids possess an origin of replication (ori), which consists of an AT-rich region, adjacent to a number of iterons which are essential in cis for replication initiation. This mode of replication requires a plasmid-encoded replication initiation protein (Rep) and is DNA polymerase I independent. Class B and C replicons do not harbor a typical ori sequence and require host-encoded DNA poly-

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“…In some related plasmid replicons, the oni has been further localized to regions that contain only the 22-bp iterons and the 10-bp direct repeats (Frere et al, 1993;Kiewiet et al, 1993a;Pedersen et al, 1994). It is now believed that the Rep proteins in these plasmids bind to their 22-bp iterons to initiate replications (Kiewiet et al, 1993a;Seegers et al, 1994). Another notable feature is the presence of two inverted repeats (IR1 and IR2) which span the putative promoter site in the FG2 plasmid replicon.…”

Section: Discussionmentioning

confidence: 99%

“…lactis FG2. To date, three lactococcal plasmid replicons, pUCL22 (Frere et al, 1993), pWV02 (Kiewiet et al, 1993a) and pCT1138 (Pedersen et al, 1994), have been characterized extensively and shown to replicate in L. lactis by the theta-replicating mechanism. Other members of this family are all closely related.…”

Section: Discussionmentioning

confidence: 99%

Cloning and sequence analysis of a plasmid replicon from Lactococcus lactis subsp. cremoris FG2.

Liu

Duan

Dunn

1997

J. Gen. Appl. Microbiol.

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A replication region from one of the Lactococcus lactis subsp. cremoris FG2 plasmids was isolated by cloning of a 4.8-kb Xbal fragment into a replication probe vector and transformation into L. lactis LM0230. A 1.8-kb region within this fragment was sequenced and confirmed by PCR subcloning to encode a functional replicon in LM0230. The replicon consists of an open reading frame encoding a putative replication protein (Rep) of 386 amino acids and a non-coding region (on) which features several structural motifs typical of other known replication origins, including a 22-bp iteron sequence tandemly repeated three and a half times, a 10-bp direct repeat and two sets of inverted repeats. The oni region could drive replication of its plasmid when supplied with the replication region in-trans. The lack of detectable single-stranded DNA during replication and the existence of extensive homology with other known lactococcal theta replicons strongly suggest that this region encodes a theta-replicating mechanism.

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Theta replication of the lactococcal plasmid pWVO2

Cited by 84 publications

References 40 publications

Structure-Function Analysis of PrsA Reveals Roles for the Parvulin-like and Flanking N- and C-terminal Domains in Protein Folding and Secretion in Bacillus subtilis

Structure-Function Analysis of PrsA Reveals Roles for the Parvulin-like and Flanking N- and C-terminal Domains in Protein Folding and Secretion in Bacillus subtilis

Identification and Characterization of an Active Plasmid Partition Mechanism for the Novel Lactococcus lactis Plasmid pCI2000

Cloning and sequence analysis of a plasmid replicon from Lactococcus lactis subsp. cremoris FG2.

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