The identification of DNA binding factors specific for as-1-like sequences in auxin-responsive regions of parA, parB and parC.

We have identified the auxin-responsive region (Aux-RR) of the parA promoter; it is derived from a gene that is induced by auxin in tobacco mesophyll protoplasts. By analyses of gain-of-function and point mutations in transgenic tobacco plants, we showed that an as-1-like sequence was required, but not alone sufficient, for auxin responsiveness of the parA promoter, as has been also shown for the parC promoter in a recent study. A gel mobility shift assay revealed that the as-1-like sequence of the parA promoter bound specifically to the as-1 binding nuclear factor, ASF-1, while the as-1-like sequence of the parC promoter bound specifically to a novel nuclear factor named ALF-1 (as-1-like sequence binding factor-1). Since these bindings were correlated with auxin-induced activation of transcription, those results suggest that ALF-1 is a new nuclear factor involved in auxin responsiveness and that it is distinct from ASF-1. Furthermore, we found that an as-1-like sequence in the AuxRR of parB bound specifically to a tobacco nuclear factor, named ALF-2, which differed from ASF-1 and ALF-1. These results suggest that the auxin responsiveness of the auxin-inducible genes parA, parB and parC are regulated by different mechanisms, even though the cis-acting elements look similar.