A programmed site-specific DNA rearrangement in Tetrahymena thermophila requires flanking polypurine tracts

Godiska, Ronald; Yao, Meng-Chao

doi:10.1016/0092-8674(90)90688-b

Cited by 96 publications

(110 citation statements)

References 28 publications

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“…2) changes made at M3, as aberrant deletions were also observed using unaltered M-elements (Godiska and Yao 1990). In some cases, smearing bands were observed.…”

Section: Asg S Specifies Deletion Boundaries At a Distancementioning

confidence: 99%

“…We have shown previously that following its introduction into developing cells, the M1 or M2 terminus of an unaltered M-element in the rDNA vector is spliced to M3, accurately reproducing the normal process to eliminate 0.9 or 0.6 kb of the internal region (Godiska and Yao 1990). Further analysis revealed that the sequence AsGs, which is located in both flanking regions of the M-element, is essential for deletion to occur.…”

Section: Asg S Specifies Deletion Boundaries At a Distancementioning

confidence: 99%

“…A cloned copy of the M-element is able to undergo rearrangement efficiently and accurately when tested by this method (Godiska and Yao 1990). This rearrangement is dependent on certain sequences that normally flank the element.…”

mentioning

confidence: 99%

“…Thus, the AsGs sequence must play an important role in DNA deletion. The 0.6-kb deletion occurs normally in all of these cases (Godiska and Yao 1990).…”

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confidence: 99%

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A distant 10-bp sequence specifies the boundaries of a programmed DNA deletion in Tetrahymena.

Godiska

James²,

Yao³

1993

Genes & Development

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Programmed DNA deletion occurs at thousands of specific sites in most ciliates studied. To understand the mechanism of this prominent DNA rearranging process, we analyzed one of the deletion elements {the M-element) in Tetrahymena by making specific mutations in cloned DNAs and testing their effects on rearrangement in vivo. We found that a 10-bp polypurine sequence {5'-AAAAAGGGGG1 plays a crucial role. This sequence is located at a short distance {-45 bpl outside of the element on either side. Removal of it abolishes the deletion process. Moving it short distances away causes the deletion boundary to move with it. Insertion of this sequence into a site within the element induces new boundaries to form near the insertion site. Sequence analysis reveals that all new boundaries created are 41-54 bp away from the sequence. Thus, this sequence is necessary and sufficient to determine the boundaries of DNA deletion, and it does so from a short distance outside of the element. These results offer a possible explanation for the control of DNA deletion in ciliates and suggest that a new type of mechanism for site-specific DNA rearrangements is involved.

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“…2) changes made at M3, as aberrant deletions were also observed using unaltered M-elements (Godiska and Yao 1990). In some cases, smearing bands were observed.…”

Section: Asg S Specifies Deletion Boundaries At a Distancementioning

confidence: 99%

Section: Asg S Specifies Deletion Boundaries At a Distancementioning

confidence: 99%

mentioning

confidence: 99%

“…Thus, the AsGs sequence must play an important role in DNA deletion. The 0.6-kb deletion occurs normally in all of these cases (Godiska and Yao 1990).…”

mentioning

confidence: 99%

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A distant 10-bp sequence specifies the boundaries of a programmed DNA deletion in Tetrahymena.

Godiska

James²,

Yao³

1993

Genes & Development

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“…An additional linker sequence (5Ј-AACC -TCGAGTGAATGATTTGAGGGCCC-3Ј) was added to the last codon of GFP that contains one leucine codon attached to an in-frame XhoI site that is followed by three out-of-frame stop codons and terminating with an ApaI site that allows fusion to coding sequences or random cDNAs. This entire rpL29-GFP cassette was sequentially inserted into the HindIII site of pHSS6 and then as a NotI fragment into rRNA gene transformation vector pD5H8 (Godiska and Yao, 1990) to create pVGF-1 that allows GFP expression in growing cells. The rpL29 promoter was then displaced with a 0.85 kbp fragment of the PDD1 promoter inserted into the PmeI site at the 5Ј end of GFP (the fusion point was nucleotide 215 of the published PDD1 sequence, Acc.…”

Section: Generation Of Gfp Expression Vectors and A Gfp-cdna Librarymentioning

confidence: 99%

Identification of novel chromatin-associated proteins involved in programmed genome rearrangements inTetrahymena

Meng

Yao

Halasz

et al. 2007

Journal of Cell Science

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Extensive DNA rearrangements occur during the differentiation of the developing somatic macronuclear genome from the germ line micronuclear genome of Tetrahymena thermophila. To identify genes encoding proteins likely to be involved in this process, we devised a cytological screen to find proteins that specifically localize in macronuclear anlagen (Lia proteins) at the stage when rearrangements occur. We compared the localization of these with that of the chromodomain protein, Pdd1p, which is the most abundant known participant in this genome reorganization. We show that in live cells, Pdd1p exhibits dynamic localization, apparently shuttling from the parental to the developing nuclei through cytoplasmic bodies called conjusomes. Visualization of GFP-tagged Pdd1p also highlights the substantial three-dimensional nuclear reorganization in the formation of nuclear foci that occur coincident with DNA rearrangements. We found that late in macronuclear differentiation, four of the newly identified proteins are organized into nuclear foci that also contain Pdd1p. These Lia proteins are encoded by primarily novel genes expressed at the beginning of macronuclear differentiation and have properties or recognizable domains that implicate them in chromatin or nucleic acid binding. Three of the Lia proteins also localize to conjusomes, a result that further implicates this structure in the regulation of DNA rearrangement.

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A micronucleus‐limited sequence family in Tetrahymena thermophila: Organization and sequence conservation

Tsao

Pearlman

1992

Dev. Genet.

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During macronuclear development in the ciliated protozoan Tetrahymena thermophila, sequence reorganization including sequence loss occurs. Addressing questions about the organization and nucleotide sequence of micronucleus limited regions can lead to insights about mechanisms of DNA rearrangements during macronuclear development as well as mechanisms for the maintenance of the stability of micronucleus-limited sequence families. We have previously identified a moderately repetitive micronucleus-limited sequence family called X-H (family members hybridize to an approximately 450 bp Xbal-HindIII restriction fragment), completely absent from macronuclear DNA. The first member of this family which we isolated is associated with terminal sequences characteristic of a Tel-1 element, a putative micronuclear transposable element. Two additional family members have been isolated which are not closely associated with Tel-1 terminal sequences. We have nucleotide sequence data for three cloned members of the X-H family. This analysis has demonstrated that the longest cloned members of the X-H family share a region of homology of approximately 2,400 bp and are highly conserved, differing only by small insertions or deletions of 100 bp or less. The sequences from one of the sequenced family members flanking the region of homology are themselves mostly micronucleus-limited.

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A programmed site-specific DNA rearrangement in Tetrahymena thermophila requires flanking polypurine tracts

Cited by 96 publications

References 28 publications

A distant 10-bp sequence specifies the boundaries of a programmed DNA deletion in Tetrahymena.

A distant 10-bp sequence specifies the boundaries of a programmed DNA deletion in Tetrahymena.

Identification of novel chromatin-associated proteins involved in programmed genome rearrangements inTetrahymena

A micronucleus‐limited sequence family in Tetrahymena thermophila: Organization and sequence conservation

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