LC8 dynein light chain (DYNLL1) binds to the C-terminal domain of ATM-interacting protein (ATMIN/ASCIZ) and regulates its subcellular localization

Journal Article · Fri Oct 28 00:00:00 EDT 2011 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/J.BBRC.2011.09.093· OSTI ID:22207537

Rapali, Peter ^[1]; Garcia-Mayoral, Maria Flor ^[2]; Martinez-Moreno, Monica ^[3]; Tarnok, Krisztian; Schlett, Katalin ^[4]; Albar, Juan Pablo ^[5]; Bruix, Marta ^[2]; Nyitray, Laszlo ^[1]; Rodriguez-Crespo, Ignacio ^[3]

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1. Dept. Biochemistry, Eoetvoes Lorand University, Budapest (Hungary)
2. Dept. Biological Physical Chemistry, IQFR, CSIC, Madrid (Spain)
3. Dept. Biochemistry and Molecular Biology I, Universidad Complutense, Madrid (Spain)
4. Dept. Physiology and Neurobiology, Eoetvoes Lorand University, Budapest (Hungary)
5. Proteomics Facility, CNB, CSIC, Madrid (Spain)

Highlights: Black-Right-Pointing-Pointer We have screened a human library with dynein light chain DYNLL1 (DLC8) as bait. Black-Right-Pointing-Pointer Dynein light chain DYNLL1 binds to ATM-kinase interacting protein (ATMIN). Black-Right-Pointing-Pointer ATMIN has 17 SQ/TQ motifs, a motif frequently found in DYNLL1-binding partners. Black-Right-Pointing-Pointer The two proteins interact in vitro, with ATMIN displaying at least five binding sites. Black-Right-Pointing-Pointer The interaction of ATMIN and DYNNL1 in transfected cells can also be observed. -- Abstract: LC8 dynein light chain (now termed DYNLL1 and DYNLL2 in mammals), a dimeric 89 amino acid protein, is a component of the dynein multi-protein complex. However a substantial amount of DYNLL1 is not associated to microtubules and it can thus interact with dozens of cellular and viral proteins that display well-defined, short linear motifs. Using DYNLL1 as bait in a yeast two-hybrid screen of a human heart library we identified ATMIN, an ATM kinase-interacting protein, as a DYNLL1-binding partner. Interestingly, ATMIN displays at least 18 SQ/TQ motifs in its sequence and DYNLL1 is known to bind to proteins with KXTQT motifs. Using pepscan and yeast two-hybrid techniques we show that DYNLL1 binds to multiple SQ/TQ motifs present in the carboxy-terminal domain of ATMIN. Recombinant expression and purification of the DYNLL1-binding region of ATMIN allowed us to obtain a polypeptide with an apparent molecular mass in gel filtration close to 400 kDa that could bind to DYNLL1 in vitro. The NMR data-driven modelled complexes of DYNLL1 with two selected ATMIN peptides revealed a similar mode of binding to that observed between DYNLL1 and other peptide targets. Remarkably, co-expression of mCherry-DYNLL1 and GFP-ATMIN mutually affected intracellular protein localization. In GFP-ATMIN expressing-cells DNA damage induced efficiently nuclear foci formation, which was partly impeded by the presence of mCherry-DYNLL1. Thus, our results imply a potential cellular interference between DYNLL1 and ATMIN functions.

• OSTI ID: 22207537
• Journal Information: Biochemical and Biophysical Research Communications, Vol. 414, Issue 3; Other Information: Copyright (c) 2011 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
AMINO ACIDS
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HEART
IN VITRO
MAMMALS
MICROTUBULES
NUCLEAR MAGNETIC RESONANCE
POLYPEPTIDES
YEASTS