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TC1320EOATR数据手册
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1
SCIENTIFIC RepoRts | 5:10243 | DOI: 10.1038/srep10243
www.nature.com/scientificreports
Chromodomain protein Tcd1 is
required for macronuclear genome
rearrangement and repair in
Tetrahymena
Jing Xu
1,2
, Yajing Yuan
2
, Aihua Liang
2
& Wei Wang
2
The survival of an organism’s progeny depends on the maintenance of its genome. Programmed DNA
rearrangement and repair in Tetrahymena occur during the dierentiation of the developing somatic
macronuclear genome from the germ line micronuclear genome. Tetrahymena chromodomain
protein (Tcd1) exhibited dynamic localization from the parental to the developing macronuclei. In the
developing macronuclei, Tcd1 colocalized with Pdd1 and H3K9me3. Furthermore, Tcd1 colocalized
with Pdd1 in the conjusome and “donut structure” of DNA elimination heterochromatin region.
During the growth and conjugation stages, TCD1 knockout cells appeared normal and similar to
wild-type strains. In addition, these knockout cells proceeded to the 2MAC-1MIC stage. However, the
progeny of the TCD1 knockout cells did not grow upon return to SPP medium and eventually died.
The deletion of the internal elimination sequence R element was partially disrupted in the developing
new macronuclei. Gamma H2A staining showed that Tcd1 loss induced the accumulation of DNA
double-strand breaks and the failure of genome repair. These results suggest that the chromodomain
protein Tcd1 is required for the rearrangement and repair of new macronuclear genome in
Tetrahymena.
e genome integrity of dierent organisms is continuously challenged by multiple internal signals and
external factors. In general, programmed DNA rearrangement and repair occur on a genome-wide scale
in ciliated protozoa. Some ciliate cells excise thousands of DNA elements from the genome, break the
chromosomes into hundreds of small units, and extensively remodel the genome during sexual develop-
ment
1
. erefore, the organisms have evolved specic molecular mechanisms to precisely and eciently
repair the broken ends generated by these events
2
. Among these protozoans, Tetrahymena is an impor-
tant and useful modern organism for exploring the molecular mechanism underlying genome rearrange-
ment and repair. Tetrahymena has two structurally and functionally dierent nuclei during the vegetative
growing stage
3
. e macronucleus (MAC) is transcriptionally active and provides the somatic functions
of cells, whereas the micronucleus (MIC) is transcriptionally silent during the vegetative growth
4
. e
germline MIC and the somatic MAC are derived from the same zygotic nucleus during Tetrahymena
sexual conjugation stage. Two types of genome rearrangement occur during new MAC formation. e
rst genome rearrangement is the deletion of internal eliminated sequences (IESs), which is accompa-
nied by the ligation of the anking macronucleus-destined sequences
5
. e other rearrangement involves
chromosomal breakage followed by the deletions of breakage eliminated sequences and the addition of
telomeres. Programmed genome rearrangements eliminate ~15% of the genome during conjugation
6
.
Subsequently, the developing new macronuclear chromosomes endoduplicate to ~45 times of the haploid
genome. In the absence of consensus DNA sequences in the process, IESs are targeted for elimination by
1
College of Life Science, Shanxi University, Taiyuan, 030006, China.
2
Key Laboratory of Chemical Biology and
Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, 030006,
China. Correspondence and requests for materials should be addressed to W.W. (email: gene@sxu.edu.cn)
Received: 18 February 2015
Accepted: 07 April 2015
Published: 19 May 2015
OPEN
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