The primary structure of rat brain (cytoplasmic) dynein heavy chain, a cytoplasmic motor enzyme.

Abstract

Overlapping cDNA clones encoding the heavy chain of rat brain cytoplasmic dynein have been isolated. The isolated cDNA clones contain an open reading frame of 13,932 bp encoding 4644 aa (Mr, 532,213). The deduced protein sequence of the heavy chain of rat brain dynein shows significant similarity to sea urchin flageilar -dynein (27.0% identical) and toDictyostelium cytoplasmic dynein (53.5% identical) throughout the entire sequence. The heavy chain of rat brain (cytoplasmic) dynein contains four putative nudeotide-binding consensus sequences [GX4GK(T/S)] in the central one-third region that are highly similar to those of sea urchin and Dictyostelium dyneins. The N-terminal one-third of the heavy chain of rat brain (cytoplasmic) dynein shows high similarity (43.8% identical) to that of Dictyostelium cytoplasmic dynein but poor similarity (19.4% identical) to that of sea urchin flageliar dynein. These results suggested that the C-terminal two-thirds of the dynein molecule is conserved and plays an essential role in microtubule-dependent motility activity, whereas the N-terminal regions are different between cytoplasmic and flageilar dyneins.The nerve cell develops a polarized morphology composed of highly branched dendrites, a long axon, and synapses. Because of the lack of protein synthesis machinery in the axon, proteins in the axon and synapses must be transported down the axon after being synthesized in the cell body. Many proteins are transported bidirectionally in membranous organelles of various kinds by fast axonal flow (200-400 mm/ day), whereas other proteins such as cytoskeletal components are conveyed by slow axonal flow (0.5-2 mm/day). Electron microscopic studies of the neuronal cytoskeleton in vivo have suggested that microtubules and crossbridges between microtubules and membranous organelles form the structural basis for fast flow (1-3). Recently, two microtubule-activated ATPases, kinesin and brain dynein [cytoplasmic dynein, microtubule-associated protein (MAP) 1C], were identified as candidates for anterograde and retrograde molecular motors, respectively (4-7). In fact, kinesin is primarily associated with anterogradely moving membranous organelles in the axon, strongly supporting the hypothesis that kinesin is really an anterograde motor in vivo (8). Brain dynein is associated with both anterogradely and retrogradely moving membranous organelles (9), consistent with the suggestion that brain dynein is transported to the nerve terminal by fast flow and subsequently functions as a retrograde transport motor in vivo. Molecular genetic and ultrastructural approaches have dissected the molecular structure and functional domains of kinesin motors (10-12). However, detailed analysis of the primary structure of brain dynein hasThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. not yet been accomplished. To further elucidate the functi...

The dynein genes of Paramecium tetraurelia: the structure and expression of the ciliary beta and cytoplasmic heavy chains.

“…There is a clear separation between the three 13 chains and the nine cytoplasmic dyneins (bold font). Dynein heavy chain sequences and their abbreviations used in this table are as follows: DHC-6, Paramecium 13 (this study; accession number U19464); SU 13, sea urchin 13 X59603); Chlamy 13, Chlamydomonas 13 (Mitchell and Brown, 1994; U02963); Chlamy -y, Chlamydomonas y (Wilkerson et al, 1994); DHC-8, Paramecium cytoplasmic (this study; U20449); Dicty, Dictyostelium (Koonce et al, 1992;Z15124); Dro cyto, Drosophila cytoplasmic (Li et al, 1994;L23195); MAPlC, rat cytoplasmic (Mikami et al, 1993;L08505); C eleg, Caenorhabditis rhabditis (Lye et al, 1995;L33260); SU cyto, sea urchin la Z21941); N crassa, Neurospora crassa (Plamann et al, 1994;L31504); Asp, Aspergillus nidulans (Xiang et al, 1994;U03904); Yeast, Saccharomyces cerevisiae (Eshel et al, 1993;Z21877 Ogawa, 1991), Chlamydomonas (Mitchell and Brown, 1994), and Paramecium (this report); the axonemal y heavy chain from Chlamydomonas (Wilkerson et al, 1994); and the cytoplasmic dyneins from Dictyostelium (Koonce et al, 1992), rat brain (Mikami et al, 1993;Zhang et al, 1993), S. cerevisiae (Eshel et al, 1993;Li et al, 1993), Aspergillus (Xiang et al, 1994), Neurospora (Plamann et al, 1994), Drosophila (Li et al, 1994), C. elegans (Lye et al, 1995), and Paramecium (this report cated by potential differences due to species and tissue variation.…”

Going mobile: microtubule motors and chromosome segregation.

“…These discoveries suggested that cytoplasmic dynein may power organelle movements and perhaps mitotic movements as well. Unlike the diverse number of kinesin-related genes, thus far there appears to be but one cytoplasmic dynein heavy chain gene in most organisms (37)(38)(39)(40)(41)(42). Recent evidence suggests that cytoplasmic dynein functional diversity is achieved through the association of the heavy chain with a variety of accessory proteins that target and regulate its activity (8,43,44 (89), and epitope-tagged CIN8 or KIP1 (48,49), shows that these bimC family members are distributed along the length of spindle microtubules during mitosis.…”

“…Due in large part to the considerable size of dynein, the identification and characterization of cytoplasmic dyneins has not progressed as rapidly as the analyses of members of the kinesin superfamily. However, recent accomplishments in the field, most importantly the cloning of full-length dynein heavy chains from a number of organisms, have opened the door to the full characterization of cytoplasmic dynein and perhaps cytoplasmic dynein-related proteins (37,39,40,41,(73)(74)(75)(76)(77).…”

Isolation of several human axonemal dynein heavy chain genes: genomic structure of the catalytic site, phylogenetic analysis and chromosomal assignment

“…We analysed data released from the GenBank data base for cytoplasmic DHCs from rats [24,25], humans [26], D. discoideum [27], yeast [28,29], E. nidulans [30], N. crassa [31], P. tetraurelia [15], C. elegans [32], D. melanogaster [33] and sea urchins [17], and for axonemal DHCs from rats [18], P. tetraurelia [15], C. reinhardtii [12,13], D. melanogaster [16] and sea urchins [17]. The oligonucleotide primers for PCR were designed according to the amino acid sequences corresponding to the most conserved regions of the PI-loop domain among the cytoplasmic or axonemal DHC sequences, and to compensate for the potential mismatches between templates P-loops 400 aa The four centrally located P-loops are indicated by boxes 1^1 on a whole dynein heavy chain depicted by black bar, box 1 (hatched box) being the catalytic functional Pl-loop.…”