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Chapter 43 : Plasmids

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Abstract:

Laboratory strains of , which are derived from strain 168, are devoid of endogenous plasmids. Many plasmids from other spp. and from other genera of gram-positive bacteria, such as staphylococci and streptococci, have been introduced, studied, and used in 168. Studies conducted in as well as in natural plasmid hosts and in are presented in this chapter, as they have all contributed to our understanding of plasmids from gram-positive bacteria. RCR plasmids have been studied much more extensively than theta replicating plasmids and have been reviewed recently in depth. The chapter focuses on newly found aspects of the replication mechanism, copy control, and interaction with host functions that help explain the life cycles of these plasmids. In contrast, theta replicating plasmids have not been previously reviewed in detail and are discussed more extensively. Plasmids of the pAMβ1 family have a large host range and are auto transferable. The following observations indicate that plasmids of the pAMβ1 family use an initiation mechanism related to that of ColEl-type replicons. First, pAMβ1, pIP501, and pSM19035 do not transform A mutant of . Second, DNA polymerase 1 initiates pAMβ1 leading-strand synthesis. Third, pAMβ1, pIP501, and pSM19035 carry a -like sequence on the lagging-strand template, ~100 bp downstream of the initiation on site of the leading-strand synthesis.

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43

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Figures

Image of Figure 1
Figure 1

Unidirectional replication of rolling-circle repli-cons. Heavy lines, plus strands; light lines, minus strands; continuous lines, parental DNA; discontinuous lines, newly synthesized DNA. “Plus origin” represents the terminus-plus-origin overlapping region, and the bent arrow indicates direction of replication. <, secondary structure at the minus origin. Steps are as follows, (i) Initiation. Nick by Rep at the plus origin. Copy number control is exerted at this step, (ii) Displacement of the plus strand and elongation from die 3′ OH nick to synthesize a new plus strand, (iii) Termination of plus-strand synthesis. Rep renicks at the plus origin to release one complete double-stranded plasmid that is then free to reenter the replication pool. Or, (iii) if termination does not occur because of, for example, plus-strand breakage or Rep protein release, high-molecular-weight linear concatemers (HMW) will form, (iv) Religation of the displaced plus strand to form a free ssDNA replication intermediate. (v) Conversion synthesis of ssDNA to dsDNA. Initiation at the minus origin (utilizing host factors) is followed by elongation to synthesize a new minus strand, (vi) Termination of conversion synthesis, releasing a second double-stranded molecule for reentry into the replication pool.

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43
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Image of Figure 2
Figure 2

Regions required for copy control, replication, and multimer resolution in three plasmids from the pAM1 family. Continuous line, sequenced regions; broken line, unsequenced regions; boxes, ORFs; arrows within the boxes, direction of transcription. The region lacking in pIP501 and pSM 19035 is shown in brackets. Replication origin and direction of replication are indicated by heavy arrows. Several restriction sites that suggest sequence homology outside of the common sequenced regions are represented. Rep and Res stand for the replication protein and resolvase, respectively. A, I; Hd, dIII; K, IHp, I; P, I; E, RI.

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43
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Image of Figure 3
Figure 3

Single-strand initiation (A) site of pAM1. The sequence of the active strand is shown. Endpoints of deletions generated by exonuclease III are indicated by arrows; + and − refer to deletions that preserve or abolish activity.

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43
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Image of Figure 4
Figure 4

Copy control regions of three plasmids of the pAMβ1 family and putative control mechanisms. (A) Critical promoters are represented by bent arrows, which indicate directions of transcription; inverted repeats are labeled I to IV. The region diverging to almost 50% between pAMβ1 and two other plasmids is boxed. Other symbols are explained in the legend to Fig. 2 . (B) Hypothetical secondary structures generated upon interaction of mRNA and countertranscript (top) and folding of the 5′ end of the mRNA (bottom).

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43
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Image of Figure 5
Figure 5

Structure of the resolution sites of pAMβ1, pIP501, and pSM 19035. The three imperfect inverted repeats (I through III) are represented by arrows, matching regions are indicated by a continuous line, and nonhomologous regions are indicated by a broken line. The numbers refer to lengths (in base pairs) of different regions and distances between the repeats. The sequence shown beneath the box is present in the nine repeats from the three plasmids.

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43
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Image of Figure 6
Figure 6

Comparison of conserved C-terminal domains of INT recombinases and Resβ and ResIP proteins. The amino acid sequences are from previously published data ( ). Alignments are based on those reported previously ( ). Positions within the INT recombinases, where at least 11 residues are related, are shown by uppercase letters (related amino acids are considered to be ST, ILMV, DE, QN, FY, and C; 50). Positions at which at least 11 residues are identical are indicated by boldface uppercase letters. Resβ and ResIP amino acids matching the consensus are shown. The arrow indicates the tyrosine likely to be transiently linked to DNA during recombination is close to the C-terminal part of the conserved domain.

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43
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Tables

Generic image for table
Table 1

Rolling-circle plasmids and their hosts of origin

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43
Generic image for table
Table 2

pAMβ1 plasmid family

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43
Generic image for table
Table 3

replicating plasmids which do not belong to the pAM1 family

Citation: Jannière L, Grass A, Ehrlich S. 1993. Plasmids, p 625-644. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch43

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