PDIP38 is a novel adaptor-like modulator of the mitochondrial AAA+ protease CLPXP

Strack, P. R.; Brodie, E. J.; Zhan, H.; Schuenemann, V. J.; Valente, L. J.; Saiyed, T.; Zeth, K.; Truscott, K. N.; Dougan, D. A.

doi:10.1101/2020.05.19.105320

bioRxiv · 10.1101/2020.05.19.105320

PDIP38 is a novel adaptor-like modulator of the mitochondrial AAA+ protease CLPXP

Strack, P. R.·Brodie, E. J.·Zhan, H.·Schuenemann, V. J.·Valente, L. J.·Saiyed, T.·Zeth, K.·Truscott, K. N.·Dougan, D. A.

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Abstract

Polymerase {delta} interacting protein of 38 kDa (PDIP38) was originally identified in a yeast two hybrid screen as an interacting protein of DNA polymerase delta, more than a decade ago. Since this time several subcellular locations have been reported and hence its function remains controversial. Our current understanding of PDIP38 function has also been hampered by a lack of detailed biochemical or structural analysis of this protein. Here we show, that human PDIP38 is directed to the mitochondrion, where it resides in the matrix compartment, together with its partner protein CLPX. PDIP38 is a bifunctional protein, composed of two conserved domains separated by an -helical hinge region (or middle domain). The N-terminal (YccV-like) domain of PDIP38 forms an SH3-like {beta}-barrel, which interacts specifically with CLPX, via the adaptor docking loop within the N-terminal Zinc binding domain (ZBD) of CLPX. In contrast, the C-terminal (DUF525) domain forms an Immunoglobin-like {beta}-sandwich fold, which contains a highly conserved hydrophobic groove. Based on the physicochemical properties of this groove, we propose that PDIP38 is required for the recognition (and delivery to CLPXP) of proteins bearing specific hydrophobic degrons, potentially located at the termini of the target protein. Significantly, interaction with PDIP38 stabilizes the steady state levels of CLPX in vivo. Consistent with these data, PDIP38 inhibits the LONM-mediated turnover of CLPX in vitro. Collectively, our findings shed new light on the mechanistic and functional significance of PDIP38, indicating that in contrast to its initial identification as a nuclear protein, PIDP38 is a bona fide mitochondrial adaptor protein for the CLPXP protease.

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Strack, P. R., Brodie, E. J., Zhan, H., Schuenemann, V. J., Valente, L. J., Saiyed, T., Zeth, K., Truscott, K. N., Dougan, D. A.. 2020-05-22. PDIP38 is a novel adaptor-like modulator of the mitochondrial AAA+ protease CLPXP. https://doi.org/10.1101/2020.05.19.105320

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