ISOLATION AND CHARACTERIZATION OF THE FOUR ARABIDOPSIS THALIANA POLY(A) POLYMERASE GENES

• Main Author: Meeks, Lisa Renee
• Format: Others
• Published: UKnowledge 2005
• Subjects: Poly(A) Polymerase|mRNA 3'-End Formation|RNA-Processing|Poly(A) Tail|Arabidopsis thaliana
• Online Access: http://uknowledge.uky.edu/gradschool_diss/447; http://uknowledge.uky.edu/cgi/viewcontent.cgi?article=1450&context=gradschool_diss

Poly(A) tail addition to pre-mRNAs is a highly coordinated and essential step in mRNA maturation involving multiple cis- and trans-acting factors. The trans-acting factor, poly(A) polymerase (PAP) plays an essential role in the polyadenylation of mRNA precursors. The Arabidopsis thaliana genome contains four putative PAP genes. We have found, using in silico analysis and transgenic plants expressing GUS under the control of the four PAP promoters, that each of these genes is expressed in overlapping, yet unique patterns. This gives rise to the possibility that these genes are not redundant and may be essential for plant survival. To further test this, inducible RNAi and T-DNA mutagenized plants were obtained and analyzed. Plants lacking all, or most, of each PAP gene product, due to RNAi induction, were not viable at any of the stages of plant growth tested. Furthermore, T-DNA PCR analysis determined that no plants containing a homozygous mutation, were viable. This data reveals that lack of any of the four PAP gene products has a significant effect on the plants ability of survive, thus indicating that each PAP gene is essential. Finally, transient expression experiments with each of the full length PAP cDNAs fused to GFP showed that the PAP I, PAP II and PAP IV gene products are localized throughout the nucleus and within nuclear speckles. The cellular localization of PAP III could not be determined.