Dynein light chain, Tctex-type 3 (DYNLT3), is a member of the cytoplasmic dynein DYNLT light chain family and has been reported to have a potential role in chromosome congression in human mitosis. However, its role in mammalian meiosis is unclear. In this study, we examined its localization, expression, and functions in mouse oocyte meiosis. Immunofluorescent staining showed that DYNLT3 was restricted to the germinal vesicle and associated with kinetochores at the germinal vesicle breakdown stage, metaphase I and metaphase II. The expression level of DYNLT3 was similar at all meiotic stages. Depletion of DYNLT3 by antibody injection resulted in chromosome misalignment and decrease of the polar body extrusion rate. We further found that DYNLT3-depleted oocytes displayed kinetochore-microtubule detachments. Chromosome-spread experiments showed that depletion of DYNLT3 inhibited the metaphase-anaphase transition by preventing homologous chromosome segregation in meiosis I. Our data suggest that DYNLT3 is required for chromosome alignment and homologous chromosome segregation during mouse oocyte meiosis.
YinSSunXFSchattenHSunQY. Molecular insights into mechanisms regulating faithful chromosome separation in female meiosis. Cell Cycle. 2008;7(19):2997–3005.
2.
MatsuuraSItoETauchiHKomatsuKIkeuchiTKajiiT. Chromosomal instability syndrome of total premature chromatid separation with mosaic variegated aneuploidy is defective in mitotic-spindle checkpoint. Am J Hum Genet. 2000;67(2):483–486.
3.
BaderJRVaughanKT. Dynein at the kinetochore: timing, interactions and functions. Semin Cell Dev Biol. 2010;21(3):269–275.
4.
VaisbergEAKoonceMPMcIntoshJR. Cytoplasmic dynein plays a role in mammalian mitotic spindle formation. J Cell Biol. 1993;123(4):849–858.
5.
BussonSDujardinDMoreauADompierreJDe MeyJR. Dynein and dynactin are localized to astral microtubules and at cortical sites in mitotic epithelial cells. Curr Biol. 1998;8(9):541–544.
6.
O'ConnellCBWangYL. Mammalian spindle orientation and position respond to changes in cell shape in a dynein-dependent fashion. Mol Biol Cell. 2000;11(5):1765–1774.
7.
PfarrCMCoueMGrissomPMHaysTSPorterMEMcIntoshJR. Cytoplasmic dynein is localized to kinetochores during mitosis. Nature. 1990;345(6272):263–265.
8.
WordemanLSteuerERSheetzMPMitchisonT. Chemical subdomains within the kinetochore domain of isolated CHO mitotic chromosomes. J Cell Biol. 1991;114(2):285–294.
9.
YangZTuluUSWadsworthPRiederCL. Kinetochore dynein is required for chromosome motion and congression independent of the spindle checkpoint. Curr Biol. 2007;17(11):973–980.
10.
WojcikEBastoRSerrMScaerouFKaressRHaysT. Kinetochore dynein: its dynamics and role in the transport of the Rough deal checkpoint protein. Nat Cell Biol. 2001;3(11):1001–1007.
11.
VorozhkoVVEmanueleMJKallioMJStukenbergPTGorbskyGJ. Multiple mechanisms of chromosome movement in vertebrate cells mediated through the Ndc80 complex and dynein/dynactin. Chromosoma. 2008;117(2):169–179.
12.
VarmaDMonzoPStehmanSAValleeRB. Direct role of dynein motor in stable kinetochore-microtubule attachment, orientation, and alignment. J Cell Biol. 2008;182(6):1045–1054.
13.
GeeMValleeR. The role of the dynein stalk in cytoplasmic and flagellar motility. Eur Biophys J. 1998;27(5):466–473.
14.
OiwaKSakakibaraH. Recent progress in dynein structure and mechanism. Curr Opin Cell Biol. 2005;17(1):98–103.
15.
ValleeRBWilliamsJCVarmaDBarnhartLE. Dynein: an ancient motor protein involved in multiple modes of transport. J Neurobiol. 2004;58(2):189–200.
16.
HaburaATikhonenkoIChisholmRLKoonceMP. Interaction mapping of a dynein heavy chain. Identification of dimerization and intermediate-chain binding domains. J Biol Chem. 1999;274(22):15447–15453.
17.
TynanSHGeeMAValleeRB. Distinct but overlapping sites within the cytoplasmic dynein heavy chain for dimerization and for intermediate chain and light intermediate chain binding. J Biol Chem. 2000;275(42):32769–32774.
18.
KingSMBarbareseEDillmanJF,3rd. Cytoplasmic dynein contains a family of differentially expressed light chains. Biochemistry. 1998;37(43):15033–15041.
19.
LaderEHaHSO'NeillMArtztKBennettD. tctex-1: a candidate gene family for a mouse t complex sterility locus. Cell. 1989;58(5):969–979.
20.
CaggeseCMoschettiRRagoneGBarsantiPCaizziR. dtctex-1, the Drosophila melanogaster homolog of a putative murine t-complex distorter encoding a dynein light chain, is required for production of functional sperm. Mol Genet Genomics. 2001;265(3):436–444.
21.
MikiFOkazakiKShimanukiMYamamotoAHiraokaYNiwaO. The 14-kDa dynein light chain-family protein Dlc1 is required for regular oscillatory nuclear movement and efficient recombination during meiotic prophase in fission yeast. Mol Biol Cell. 2002;13(3):930–946.
TsengYYGruzdevaNLiAChuangJZSungCH. Identification of the Tctex-1 regulatory element that directs expression to neural stem/progenitor cells in developing and adult brain. J Comp Neurol. 2010;518(16):3327–3342.
24.
KaiNMishinaMYagiT. Molecular cloning of Fyn-associated molecules in the mouse central nervous system. J Neurosci Res. 1997;48(5):407–424.
25.
NaganoFOritaSSasakiT. Interaction of Doc2 with tctex-1, a light chain of cytoplasmic dynein. Implication in dynein-dependent vesicle transport. J Biol Chem. 1998;273(46):30065–30068.
26.
BauchACampbellKSRethM. Interaction of the CD5 cytoplasmic domain with the Ca2+/calmodulin-dependent kinase IIdelta. Eur J Immunol. 1998;28(7):2167–2177.
27.
TaiAWChuangJZBodeCWolfrumUSungCH. Rhodopsin’s carboxy-terminal cytoplasmic tail acts as a membrane receptor for cytoplasmic dynein by binding to the dynein light chain Tctex-1. Cell. 1999;97(7):877–887.
28.
MokYKLoKWZhangM. Structure of Tctex-1 and its interaction with cytoplasmic dynein intermediate chain. J Biol Chem. 2001;276(17):14067–14074.
29.
NadanoDNakayamaJMatsuzawaSSatoTAMatsudaTFukudaMN. Human tastin, a proline-rich cytoplasmic protein, associates with the microtubular cytoskeleton. Biochem J. 2002;364(pt 3):669–677.
30.
MuellerSCaoXWelkerRWimmerE. Interaction of the poliovirus receptor CD155 with the dynein light chain Tctex-1 and its implication for poliovirus pathogenesis. J Biol Chem. 2002;277(10):7897–7904.
31.
MachadoRDRudarakanchanaNAtkinsonC. Functional interaction between BMPR-II and Tctex-1, a light chain of Dynein, is isoform-specific and disrupted by mutations underlying primary pulmonary hypertension. Hum Mol Genet. 2003;12(24):3277–3286.
32.
SugaiMSaitoMSukegawaI. PTH/PTH-related protein receptor interacts directly with Tctex-1 through its COOH terminus. Biochem Biophys Res Commun. 2003;311(1):24–31.
33.
DouglasMWDiefenbachRJHomaFL. Herpes simplex virus type 1 capsid protein VP26 interacts with dynein light chains RP3 and Tctex1 and plays a role in retrograde cellular transport. J Biol Chem. 2004;279(27):28522–28530.
34.
Gauthier-FisherALinDCGreeveMKaplanDRRottapelRMillerFD. Lfc and Tctex-1 regulate the genesis of neurons from cortical precursor cells. Nat Neurosci. 2009;12(6):735–744.
35.
ChuangJZMilnerTASungCH. Subunit heterogeneity of cytoplasmic dynein: Differential expression of 14 kDa dynein light chains in rat hippocampus. J Neurosci. 2001;21(15):5501–5512.
36.
WuHMaciejewskiMWTakebeSKingSM. Solution structure of the Tctex1 dimer reveals a mechanism for dynein-cargo interactions. Structure. 2005;13(2):213–223.
37.
SachdevPMenonSKastnerDB. G protein beta gamma subunit interaction with the dynein light-chain component Tctex-1 regulates neurite outgrowth. EMBO J. 2007;26(11):2621–2632.
38.
LoKWKogoyJMPfisterKK. The DYNLT3 light chain directly links cytoplasmic dynein to a spindle checkpoint protein, Bub3. J Biol Chem. 2007;282(15):11205–11212.
39.
ApcarianACunninghamALDiefenbachRJ. Identification of binding domains in the herpes simplex virus type 1 small capsid protein pUL35 (VP26). J Gen Virol. 2010;91(pt 11):2659–2663.
40.
DohnerKRadtkeKSchmidtSSodeikB. Eclipse phase of herpes simplex virus type 1 infection: efficient dynein-mediated capsid transport without the small capsid protein VP26. J Virol. 2006;80(16):8211–8224.
41.
TaiAWChuangJZSungCH. Localization of Tctex-1, a cytoplasmic dynein light chain, to the Golgi apparatus and evidence for dynein complex heterogeneity. J Biol Chem. 1998;273(31):19639–19649.
42.
YangSLiuXYinYFukudaMNZhouJ. Tastin is required for bipolar spindle assembly and centrosome integrity during mitosis. FASEB J. 2008;22(6):1960–1972.
43.
PaschalBMValleeRB. Retrograde transport by the microtubule-associated protein MAP 1C. Nature. 1987;330(6144):181–183.
44.
EcheverriCJPaschalBMVaughanKTValleeRB. Molecular characterization of the 50-kD subunit of dynactin reveals function for the complex in chromosome alignment and spindle organization during mitosis. J Cell Biol. 1996;132(4):617–633.
45.
WassmannKNiaultTMaroB. Metaphase I arrest upon activation of the Mad2-dependent spindle checkpoint in mouse oocytes. Curr Biol. 2003;13(18):1596–1608.
46.
LiMLiSYuanJ. Bub3 is a spindle assembly checkpoint protein regulating chromosome segregation during mouse oocyte meiosis. PLoS One. 2009;4(11):e7701.
47.
WeiLLiangXWZhangQH. BubR1 is a spindle assembly checkpoint protein regulating meiotic cell cycle progression of mouse oocyte. Cell Cycle. 2010;9(6):1112–1121.
48.
ZhangDYinSJiangMX. Cytoplasmic dynein participates in meiotic checkpoint inactivation in mouse oocytes by transporting cytoplasmic mitotic arrest-deficient (Mad) proteins from kinetochores to spindle poles. Reproduction. 2007;133(4):685–695.
