Identification of an essential core element and stimulatory sequences in a <i>Kluyveromyces lactis</i> ARS element, <i>KARS101</i>

Fabiani, Lucia; Frontali, Laura; Newlon, Carol S.

doi:10.1046/j.1365-2958.1996.401938.x

Cited by 17 publications

(44 citation statements)

References 32 publications

(56 reference statements)

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“…The ORI sequences of Y. lipolytica are not strikingly AT rich, at least within the fragments we considered in this study; this is contrary to the situation in S. cerevisiae (61,72), K. lactis (24), Hansenula polymorpha (6,68), and S. pombe (58,80). A similar situation exists in the slime mold Physarum polycephalum, for which the efficient bidirectional replication origins so far described are actually relatively GC rich, with no striking sequence similarities between the four sequenced initiation sites (4,5).…”

Section: Discussioncontrasting

confidence: 83%

“…An ARS consensus has been described for Kluyveromyces lactis (23); this consensus is also present in ARS sequences from Kluyveromyces marxianus (47). In K. lactis the consensus is dispensable for the function of a large ARS fragment but stimulates the replication efficiency of a smaller ARS fragment (24).…”

Section: Although the Nature Of Higher Eukaryotic Replication Originsmentioning

confidence: 96%

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An Origin of Replication and a Centromere Are Both Needed To Establish a Replicative Plasmid in the YeastYarrowia lipolytica

Vernis

Abbas

Chasles

et al. 1997

Molecular and Cellular Biology

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. We have used the integration properties of centromeric sequences to show that all Y. lipolytica ARS elements so far isolated are composed of both a replication origin and a centromere. The sequence and the distance between the origin and centromere do not seem to play a critical role, and many origins can function in association with one given centromere. A centromeric plasmid can therefore be used to clone putative chromosomal origins coming from several genomic locations, which confer the replicative property on the plasmid. The DNA sequences responsible for initiation in plasmids are short (several hundred base pairs) stretches which map close to or at replication initiation sites in the chromosome. Their chromosomal deletion abolishes initiation, but changing their chromosomal environment does not. Although the nature of higher eukaryotic replication originsis not yet clear (12), that of origins in the yeast Saccharomyces cerevisiae is relatively well understood. It has been possible to clone S. cerevisiae genomic sequences that confer on a plasmid the ability to replicate extrachromosomally in this yeast. These sequences are called ARS (autonomously replicating sequence) elements. Most ARS elements are active as origins in their chromosomal context, as shown by two-dimensional (2D) gel replicon mapping studies (8,44,62). ARS elements are usually smaller than 150 bp, and they contain an essential motif that matches the 11-bp ARS consensus sequence (ACS; WTT TAYRTTTW) in at least 9 of 11 positions (61, 72). The ACS is essential for initiation of replication, both on plasmids and in the chromosome (17). Also essential is domain B, located at the 3Ј end of the ACS (54,55,76). Depending on the ARS element, domain B can be subdivided into two or three subdomains with variable sequences but conserved roles (42, 66). Finally, a DNA unwinding element is frequently present in ARS elements and is important for replication of plasmids (60) and chromosomes (41). Initiation at ARS elements is under strict cell cycle control (reviewed in reference 18).In contrast to the defined origin sequences of S. cerevisiae, the degree to which specific sequences are employed as origins in the differentiated cells of higher organisms is not yet clear. Most studies employing labeling techniques suggest that initiation takes place at specific sites or in very small (a few kilobases or less) initiation zones (15,19,35). In contrast, all studies employing 2D gel replicon mapping techniques-and some studies using labeling techniques (31, 71)-suggest that initiation can take place at any of numerous locations within large initiation zones of 50 kb or more (reviewed in reference 36). Furthermore, in contrast to the discrete ARS elements of S. cerevisiae, most human DNA pieces larger than ϳ10 kb permit extrachromosomal plasmid replication (38,51). In this case, initiation does not start at a preferred locus, a result which is also observed in plasmid transformation in Xenopus eggs (46,53). Demonstrations that the choice of initiation sites ...

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Section: Discussioncontrasting

confidence: 83%

Section: Although the Nature Of Higher Eukaryotic Replication Originsmentioning

confidence: 96%

An Origin of Replication and a Centromere Are Both Needed To Establish a Replicative Plasmid in the YeastYarrowia lipolytica

Vernis

Abbas

Chasles

et al. 1997

Molecular and Cellular Biology

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“…Our previous analysis of a K. lactis chromosomal DNA fragment with ARS activity in both K. lactis and S. cerevisiae, KARS101, revealed that ARS activity in K. lactis depended upon a 40-bp core sequence (Fabiani et al 1990(Fabiani et al , 1996. Alignment of the 159-bp pKD1 fragment with a 397-bp fragment carrying KARS101 revealed a 49-bp region showing 53% identity (Fig.…”

Section: Sequence Of the Pkd1 Ars Elementmentioning

confidence: 89%

“…The 103-bp region contained a 12-bp sequence (5¢-ATTTATTGTTTT-3¢) that is related to and functions as the S. cerevisiae ACS. Analysis of deletion and linker substitution mutations in the 103-bp fragment showed that the essential core for the K. lactis ARS is 40 nucleotides in length, at least twice as large as domain A in S. cerevisiae, and that dierent sequences are required for ARS function in the two organisms (Fabiani et al 1996).…”

Section: Introductionmentioning

confidence: 99%

A DNA replication origin and a replication fork barrier used in vivo in the circular plasmid pKD1

et al. 2001

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As in other yeasts, ARS-containing plasmids can be maintained extrachromosomally in Kluyveromyces lactis. Although some fragments of K. lactis DNA have ARS activity in both K. lactis and Saccharomyces cerevisiae, it appears that the sequences required for ARS activity in the two yeasts are different. As an approach to a better understanding of ARS structure and function in K. lactis, we analyzed the replication of the circular plasmid pKD1. We identified a 159-bp sequence able to promote autonomous replication of pKD1 in both yeasts; this fragments contains both a sequence related to the S. cerevisiae ARS consensus sequence and a region of 53% identity to the 40-bp sequence essential for K. lactis KARS101 function. By the analysis of in vivo replication intermediates we provide the first direct evidence that DNA replication initiates at or near the K. lactis ARS element. Replication terminates at the cisacting stability locus of pKD1, which functions as a replication fork barrier (RFB) and is necessary for proper plasmid segregation. RFB activity requires the pKDI gene products that are important for plasmid segregation, suggesting a link between DNA replication termination and plasmid segregation in a eukaryotic organism.

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“…The sizes of these four C. utilis ARSs were larger than that found in S. cerevisiae (100-150 bp), Kluyveromyces lactis (103 bp) and Candida albicans (709 bp), 20,24,25) and they were more similar to that found in Schizosaccharomyces pombe (500-1,500 bp).…”

Section: Discussionmentioning

confidence: 53%

Identification and Application of Novel Autonomously Replicating Sequences (ARSs) for Promoter-Cloning and Co-Transformation inCandida utilis

Ikushima¹,

Minato²,

Kondö³

2009

Bioscience, Biotechnology, and Biochemistry

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Identification of an essential core element and stimulatory sequences in a Kluyveromyces lactis ARS element, KARS101

Cited by 17 publications

References 32 publications

An Origin of Replication and a Centromere Are Both Needed To Establish a Replicative Plasmid in the YeastYarrowia lipolytica

An Origin of Replication and a Centromere Are Both Needed To Establish a Replicative Plasmid in the YeastYarrowia lipolytica

A DNA replication origin and a replication fork barrier used in vivo in the circular plasmid pKD1

Identification and Application of Novel Autonomously Replicating Sequences (ARSs) for Promoter-Cloning and Co-Transformation inCandida utilis

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