Two-triplet CGA repeats impede DNA replication in bacteriophage M13 in Escherichia coli

Expansion and contraction instabilities associated with CAG, CGG, GAA and CGA (GAC) repeats propagation cause more than a dozen human genetic diseases and cancers. In this work, the propagation behavior of a bacteriophage M13 carrying a calf prochymosin cDNA fragment with a (CGA)(2) repeat in a small hairpin forming region is reported. Such a M13 derivative when propagated in Escherichia coli, produces small plaques by decreasing phage yield and also mitigates the inhibition on host cell growth, compared to those control bacteriophages either containing a "CTGCTA" sequence or wildtype, suggesting that CGA(2) repeat impedes DNA replication in vivo. Moreover, an increased internal free energy is found associated with (CGA)(2) sequence compared to those "CTGCTA" and wildtype, which ruled out a possibility of CGA(2) repeat effects on propagation is through influencing the hairpin structure formation. (C) 2004 Published by Elsevier GmbH.