Transgenic expression of CBBP, a CXC domain protein, establishes paramutation in maize

Paramutation is the ability of specific DNA sequences to communicate in trans to establish meiotically heritable expression states. Paramutation at the maize b1 locus is mediated by seven unique noncoding transcribed tandem repeats of 853 bp that are required to establish and maintain the meiotically heritable expression and distinct chromatin states associated with b1 paramutation. In this study, we report the identification of a CXC-domain protein CBBP (CXC domain b1-repeat binding protein) that binds to a defined region within the b1 tandem repeat sequence in vivo and in vitro. When CBBP is expressed from a transgene in maize, it can induce a silent state at the b1 locus that is heritable in progeny no longer containing the transgene, and the silent epiallele is capable of silencing an active epiallele, characteristic of paramutation. Accumulation of the CBBP protein correlates with b1 silencing in transgenic and nontransgenic plants. The ability of CBBP to form multimers and to bind to the b1 tandem repeats suggests a model for counting the number of b1 repeats. In contrast to previously identified proteins involved in paramutation, CBBP does not share similarity to the known components of the Arabidopsis RNAi heterochromatin silencing pathway. Thus, this study defines another class of protein that is involved in heritable gene silencing.