Vg1 RNA localization in oocytes in the absence of xVICKZ3 RNA-binding activity.
Vg 1 RNA becomes localized at the vegetal cortex of Xenopus oocytes in a process requiring both intact microtubules (MT) and microfilaments. This localization occurs during a narrow window of oogenesis, when a number of RNA-binding proteins associate with the RNA. xVICKZ3 (Vg1 RBP/Vera), the first Vg1 RNA-binding protein identified, helps mediate the association of Vg1 RNA with MT and is co-localized with the RNA at the vegetal cortex. Given the complexity of the Vg1 RNA ribonucleoprotein (RNP) complex, it has remained unclear how xVICKZ3 functions in Vg1 RNA localization. Here, we have taken a closer look at the process of xVICKZ3 localization in oocytes. We have made use of deletion constructs to perform a structure-function analysis of xVICKZ3. The ability of xVICKZ3-GFP constructs to vegetally localize correlates with their association to MT but not with Vg1 RNA-binding ability. We find that when the ability of xVICKZ3 to bind Vg1 RNA is inhibited by the injection of a construct that dominantly inhibits RNA binding, both the construct and Vg1 RNA still localize, apparently through their continued association with a Vg1 RNA-containing RNP complex. These results emphasize the importance of protein-protein interactions in both xVICKZ3 and Vg1 RNA localization.
Rand
Kinneret
K
Hebrew University, Hadassah Medical School, Institute for Medical Research, Department of Anatomy and Cell Biology, Jerusalem, Israel.
Yisraeli
Joel K
JK
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D008969:Molecular Sequence Data; D009865:Oocytes / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D012313:RNA / Q000378:metabolism*; D034443:RNA Transport; D016601:RNA-Binding Proteins / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000502:physiology*; D014981:Xenopus / Q000196:embryology; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*
C513103:GDF1 protein, Xenopus; C498635:IGF2BP3 protein, Xenopus; D016601:RNA-Binding Proteins; D016212:Transforming Growth Factor beta; D029867:Xenopus Proteins; D012313:RNA