Helicase SUV3, Polynucleotide Phosphorylase PNPase, and Mitochondrial Polyadenylation Polymerase mtPAP Form a Transient Complex to Modulate Mitochondrial mRNA Polyadenylated Tail Lengths in Response to Energetic Changes [Enzymology]

April 25th, 2014 by Wang, D. D.-H., Guo, X. E., Modrek, A. S., Chen, C.-F., Chen, P.-L., Lee, W.-H.

Mammalian mitochondrial mRNA (mt-mRNA) transcripts are polyadenylated at 3' end with different lengths. The SUV3/PNPase complex and mtPAP have been shown to degrade and polyadenylate mitochondrial mRNA, respectively. How these two opposite actions are coordinated to modulate mt-mRNA poly(A) lengths is of interest to pursue. Here, we demonstrated that a fraction of the SUV3/PNPase complex interacts with mtPAP under low mitochondrial matrix inorganic phosphate (Pi) condition. In vitro binding experiments using purified proteins suggested that SUV3 binds to mtPAP through the N-terminal region around a.a. 100-104, distinctive from the C-terminal region around a.a. 510-514 of SUV3 for PNPase binding. mtPAP does not interact with PNPase directly, and SUV3 served as a bridge capable of simultaneously binding with mtPAP and PNPase. The complex consists of a SUV3 dimer, a mtPAP dimer and a PNPase trimer, based on the molecular sizing experiments. Mechanistically, SUV3 provides a robust single-strand RNA (ssRNA) binding domain to enhance the polyadenylation activity of mtPAP. Furthermore, purified SUV3/PNPase/mtPAP complex is capable of lengthening or shortening the RNA poly(A) tail lengths in low or high Pi/ATP ratios, respectively. Consistently, the poly(A) tail lengths of mt-mRNA transcripts can be lengthened or shortened by altering the mitochondrial matrix Pi levels via selective inhibition of the electron transport chain (ETC) or ATP synthase, respectively. Taken together, these results suggested that SUV3/PNPase/mtPAP form a transient complex to modulate mt-mRNA poly(A) tail lengths in response to cellular energy changes.
  • Posted in Journal of Biological Chemistry, Publications
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