A conserved role for the zinc finger polyadenosine RNA binding protein, ZC3H14, in control of poly(A) tail length

Neuronal expression of ZC3H14-iso1 rescues the poly(A) tail length defect of homozygous dNab2^null flies. (A) Total RNA was isolated from flies of the indicated genotypes (“WT dNab2”=control, “dNab2 null”=ex3/ex3, “+dNab2” or +“ZC3H14-iso1” = the corresponding protein expressed in dNab2^null neurons with elav-Gal4), end-labeled with [³²P]pCp, and non-poly(A) tracts were digested with RNAses A and T1. Labeled RNA was processed as described in Materials and Methods. The position of a 100-nt marker is indicated on the gel. (B) Bulk poly(A) tail length from fly heads of the indicated genotypes corresponding to the samples shown in the gel in A was analyzed by densitometric quantification of poly(A) tracts using ImageJ and plotted as normalized signal intensity (Relative Intensity) as a function of Poly(A) Tail Length in base pairs using Microsoft Excel. The approximate positions of the nucleotide size markers (nt) are indicated.