CLIP-170 homologue and NUDE play overlapping roles in NUDF localization in Aspergillus nidulans.
Proteins in the cytoplasmic dynein pathway accumulate at the microtubule plus end, giving the appearance of comets when observed in live cells. The targeting mechanism for NUDF (LIS1/Pac1) of Aspergillus nidulans, a key component of the dynein pathway, has not been clear. Previous studies have demonstrated physical interactions of NUDF/LIS1/Pac1 with both NUDE/NUDEL/Ndl1 and CLIP-170/Bik1. Here, we have identified the A. nidulans CLIP-170 homologue, CLIPA. The clipA deletion did not cause an obvious nuclear distribution phenotype but affected cytoplasmic microtubules in an unexpected manner. Although more microtubules failed to undergo long-range growth toward the hyphal tip at 32 degrees C, those that reached the hyphal tip were less likely to undergo catastrophe. Thus, in addition to acting as a growth-promoting factor, CLIPA also promotes microtubule dynamics. In the absence of CLIPA, green fluorescent protein-labeled cytoplasmic dynein heavy chain, p150(Glued) dynactin, and NUDF were all seen as plus-end comets at 32 degrees C. However, under the same conditions, deletion of both clipA and nudE almost completely abolished NUDF comets, although nudE deletion itself did not cause a dramatic change in NUDF localization. Based on these results, we suggest that CLIPA and NUDE both recruit NUDF to the microtubule plus end. The plus-end localization of CLIPA itself seems to be regulated by different mechanisms under different physiological conditions. Although the KipA kinesin (Kip2/Tea2 homologue) did not affect plus-end localization of CLIPA at 32 degrees C, it was required for enhancing plus-end accumulation of CLIPA at an elevated temperature (42 degrees C).
Department of Pharmacology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA.
Vladimir P
VP
Efimov
Jun
J
Zhang
Xin
X
Xiang
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D000072159:Dynactin Complex; D005656:Fungal Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D017353:Gene Deletion; D006358:Hot Temperature; D016547:Kinesins / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*
D000072159:Dynactin Complex; D005656:Fungal Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; C121041:RO-11 protein, Neurospora crassa; C076744:cytoplasmic linker protein 170; D016547:Kinesins