Vernos I et al. (1993), Multiple kinesin-like transcripts in Xenopus oo...

XB-ART-22617

Dev Biol 1993 May 01;1571:232-9. doi: 10.1006/dbio.1993.1127.

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Multiple kinesin-like transcripts in Xenopus oocytes.

Vernos I , Heasman J , Wylie C .

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Recent evidence shows that kinesin-like proteins (Klps) form a very large multigene family. A recent study using the polymerase chain reaction (PCR) identified six new candidate Klps in Drosophila, making the total number of members of this family in Drosophila at least 11 (Stewart et al., 1991, Proc. Natl. Acad. Sci. USA 88, 4424-4427). The functional basis of this diversity is not clear. Different Klps could have cell type-specific functions, or they could perform different functions within the same cell type, or a mixture of both. To investigate the degree to which different Klps are expressed in the same cell, we chose the Xenopus oocyte. During oocyte differentiation, and in the egg, different types of microtubule-based motility occur; all are important to the normal development of the embryo after fertilization. Using PCR we identified and partially sequenced four novel Klp mRNAs from the Xenopus oocyte (denoted XKlps 1-4). Multialign sequence comparison suggests that one of them, XKlp3, may be the Xenopus counterpart of Drosophila Klp4. Similarly Xenopus Eg5 is closely related to Drosophila Klp2. Northern blot analysis reveals that the Xenopus XKlps have different patterns of expression during embryogenesis. These data show that at least four Klps can exist in the same cell and that they can be differentially regulated during early development, and suggest their differential function in oogenesis and early development.

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Species referenced: Xenopus laevis
Genes referenced: kif11 kif15 kif3b kif4a stard9 tbx2

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	FIG. 1. Sequence alignment between the four new Xenopus kinesin-like molecules, the Xenopu..;; kinesin-like Eg5, and Drosophila kinesin. Residues that are homologous, or represent conservative mutations in all the sequences (I, V, L; Y, F; T, S; E, Q; E, D; N, D; K, R), are shown in bold. Underlined residues represent a nonconservative mutation in an otherwise conserved amino acid in the rest of the kinesin sequences.
	FIG. 2. Sequence comparison between XKlp3 and Drosophila Klp4, and Klp5. (A) Comparison between XKlp3 and Klp4. (B) Comparison between XKJp3 and Klp5. Res idues that are identical are indicated by two dots. Conservative mutations (T, S; D, N; E, Q; I, L, V; Y, F; K, R; D, E) are indicated by one dot.
	FIG. 3. Sequence comparison between Xenopus Eg5 and Drosophila Klp2. Residues that are identical are indicated by two dots. Conservative mutations (T, S; D, N; E, Q; I, L, V; Y, F; K, R~ D, E) are indicated by one dot.
	FIG. 4. Northern blots show the expression of the four Xenopu.;; Klps during oogenesis and early development. (A) Probed with XKlpl, then stripped and repro bed with XKlp2. The 288 band from the original gel photograph is also shown. (B) Probed with XK!p3, then stripped and reprobed with EFla, a ubiquitous gene product which codes for a translation initiation factor, and used to check for RNA loading. (C) Probed with the PCR fragmen t corresponding to XK\p4; RNA loading is shown using the photograph of the 288 RNA in the original gel. Roman numbers indicate oocyte stages and arabic numbers indicate embryonic stages.