The pathway to detangle a scrambled gene.
BACKGROUND
Programmed DNA elimination and reorganization frequently occur during cellular differentiation. Development of the somatic macronucleus in some ciliates presents an extreme case, involving excision of internal eliminated sequences (IESs) that interrupt coding DNA segments (macronuclear destined sequences, MDSs), as well as removal of transposon-like elements and extensive genome fragmentation, leading to 98% genome reduction in Stylonychia lemnae. Approximately 20-30% of the genes are estimated to be scrambled in the germline micronucleus, with coding segment order permuted and present in either orientation on micronuclear chromosomes. Massive genome rearrangements are therefore critical for development.


RESULTS
To understand the process of DNA deletion and reorganization during macronuclear development, we examined the population of DNA molecules during assembly of different scrambled genes in two related organisms in a developmental time-course by PCR. The data suggest that removal of conventional IESs usually occurs first, accompanied by a surprising level of error at this step. The complex events of inversion and translocation seem to occur after repair and excision of all conventional IESs and via multiple pathways.


CONCLUSIONS
This study reveals a temporal order of DNA rearrangements during the processing of a scrambled gene, with simpler events usually preceding more complex ones. The surprising observation of a hidden layer of errors, absent from the mature macronucleus but present during development, also underscores the need for repair or screening of incorrectly-assembled DNA molecules.
Institute of Cell Biology, University Witten/Herdecke, Witten, Germany.
Matthias
M
Möllenbeck
Yi
Y
Zhou
Andre R O
AR
Cavalcanti
Franziska
F
Jönsson
Brian P
BP
Higgins
Wei-Jen
WJ
Chang
Stefan
S
Juranek
Thomas G
TG
Doak
Grzegorz
G
Rozenberg
Hans J
HJ
Lipps
Laura F
LF
Landweber
D000818:Animals; D002467:Cell Nucleus / Q000235:genetics; D016798:Ciliophora / Q000235:genetics; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics*; D015321:Gene Rearrangement; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017384:Sequence Deletion
D016054:DNA, Protozoan