Interaction of the DYNLT (TCTEX1/RP3) light chains and the intermediate chains reveals novel intersubunit regulation during assembly of the dynein complex.
The cytoplasmic dynein 1 cargo binding domain is formed by five subunits including the intermediate chain and the DYNLT, DYNLL, and DYNLRB light chain families. Six isoforms of the intermediate chain and two isoforms of each of the light chain families have been identified in mammals. There is evidence that different subunit isoforms are involved in regulating dynein function, in particular linking dynein to different cargoes. However, it is unclear how the subunit isoforms are assembled or if there is any specificity to their interactions. Co-immunoprecipitation using DYNLT-specific antibodies reveals that dynein complexes with DYNLT light chains also contain the DYNLL and DYNLRB light chains. The DYNLT light chains, but not DYNLL light chains, associate exclusively with the dynein complex. Yeast two-hybrid and co-immunoprecipitation assays demonstrate that both members of the DYNLT family are capable of forming homodimers and heterodimers. In addition, both homodimers of the DYNLT family bind all six intermediate chain isoforms. However, DYNLT heterodimers do not bind to the intermediate chain. Thus, whereas all combinations of DYNLT light chain dimers can be made, not all of the possible combinations of the isoforms are utilized during the assembly of the dynein complex.
Lo
Kevin W-H
KW
Department of Cell Biology, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA.
Kogoy
John M
JM
Rasoul
Bareza A
BA
King
Stephen M
SM
Pfister
K Kevin
KK
D000818:Animals; D056445:Cytoplasmic Dyneins; D004398:Dyneins / Q000378:metabolism*; D006801:Humans; D046912:Multiprotein Complexes / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D051381:Rats
C504797:DYNLRB1 protein, human; D046912:Multiprotein Complexes; D020033:Protein Isoforms; C427758:DYNLL1 protein, human; C519850:DYNLT3 protein, rat; D056445:Cytoplasmic Dyneins; D004398:Dyneins