TY - JOUR
T1 - Sas-4 provides a scaffold for cytoplasmic complexes and tethers them in a centrosome
AU - Gopalakrishnan, Jayachandran
AU - Mennella, Vito
AU - Blachon, Stephanie
AU - Zhai, Bo
AU - Smith, Andrew H.
AU - Megraw, Timothy L.
AU - Nicastro, Daniela
AU - Gygi, Steven P.
AU - Agard, David A.
AU - Avidor-Reiss, Tomer
N1 - Funding Information:
We would like to thank Dr J. Iwasa for scientific illustrations, Dr F. McKeon for helping us to generate the Sas-4 monoclonal antibody, Dr T. Kaufman, Dr E. Schejter, Dr V. Corces, Dr J. Raff and Dr M. Bettencourt for providing antibodies. We are grateful to Dr T.K. Tang and Dr Glover for their gifts of a Sas-4 and Asl bacterial expression constructs. We thank Dr E. Koundakjian, Dr M. Moritz, Marcus Basiri, Frederick Yiu-Cheung Chim, and Kela Roberts for their technical help and scientific discussions. We would like to acknowledge Dr S. Ito, Dr P. Coughlin and the staff of the EM facility at Harvard Medical School for their help with our EM analyses. We thank the Nikon Imaging Center at Harvard Medical School for advice and guidance on microscopy. This work was partially supported by a grant from the Stewart Trust ‘Pilot Projects.’
PY - 2011
Y1 - 2011
N2 - Centrosomes are conserved organelles that are essential for accurate cell division and cilium formation. A centrosome consists of a pair of centrioles surrounded by a protein network of pericentriolar material (PCM) that is essential for the centrosome's function. In this study, we show that Sas-4 provides a scaffold for cytoplasmic complexes (named S-CAP), which include CNN, Asl and D-PLP, proteins that are all found in the centrosomes at the vicinity of the centriole. When Sas-4 is absent, nascent procentrioles are unstable and lack PCM, and functional centrosomes are not generated. When Sas-4 is mutated, so that it cannot form S-CAP complexes, centrosomes are present but with dramatically reduced levels of PCM. Finally, purified S-CAP complexes or recombinant Sas-4 can bind centrosomes stripped of PCM, whereas recombinant CNN or Asl cannot. In summary, PCM assembly begins in the cytosol where Sas-4 provides a scaffold for pre-assembled cytoplasmic complexes before tethering of the complexes in a centrosome.
AB - Centrosomes are conserved organelles that are essential for accurate cell division and cilium formation. A centrosome consists of a pair of centrioles surrounded by a protein network of pericentriolar material (PCM) that is essential for the centrosome's function. In this study, we show that Sas-4 provides a scaffold for cytoplasmic complexes (named S-CAP), which include CNN, Asl and D-PLP, proteins that are all found in the centrosomes at the vicinity of the centriole. When Sas-4 is absent, nascent procentrioles are unstable and lack PCM, and functional centrosomes are not generated. When Sas-4 is mutated, so that it cannot form S-CAP complexes, centrosomes are present but with dramatically reduced levels of PCM. Finally, purified S-CAP complexes or recombinant Sas-4 can bind centrosomes stripped of PCM, whereas recombinant CNN or Asl cannot. In summary, PCM assembly begins in the cytosol where Sas-4 provides a scaffold for pre-assembled cytoplasmic complexes before tethering of the complexes in a centrosome.
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U2 - 10.1038/ncomms1367
DO - 10.1038/ncomms1367
M3 - Article
C2 - 21694707
AN - SCOPUS:79959547445
SN - 2041-1723
VL - 2
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 359
ER -