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Comparative analysis of pre‐replication complex proteins in transformed and normal cells
Author(s) -
Di Paola Domenic,
ZannisHadjopoulos Maria
Publication year - 2012
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.24006
Subject(s) - biology , dna replication factor cdt1 , chromatin , origin recognition complex , licensing factor , replication factor c , pre replication complex , gene , origin of replication , dna replication , genetics , microbiology and biotechnology , control of chromosome duplication , eukaryotic dna replication
This study examines the abundance of the major protein constituents of the pre‐replication complex (pre‐RC), both genome‐wide and in association with specific replication origins, namely the lamin B2 , c‐myc , 20mer1, and 20mer2 origins. Several pre‐RC protein components, namely ORC1‐6, Cdc6, Cdt1, MCM4, MCM7, as well as additional replication proteins, such as Ku70/86, 14‐3‐3, Cdc45, and PCNA, were comparatively and quantitatively analyzed in both transformed and normal cells. The results show that these proteins are overexpressed and more abundantly bound to chromatin in the transformed compared to normal cells. Interestingly, the 20mer1, 20mer2, and c‐myc origins exhibited a two‐ to threefold greater origin activity and a two‐ to threefold greater in vivo association of the pre‐RC proteins with these origins in the transformed cells, whereas the origin associated with the housekeeping lamin B2 gene exhibited both similar levels of activity and in vivo association of these pre‐RC proteins in both cell types. Overall, the results indicate that cellular transformation is associated with an overexpression and increased chromatin association of the pre‐RC proteins. This study is significant, because it represents the most systematic comprehensive analysis done to date, using multiple replication proteins and different replication origins in both normal and transformed cell lines. J. Cell. Biochem. 113: 1333–1347, 2012. © 2011 Wiley Periodicals, Inc.