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R. S. Quatrano

Publications and source records attributed to R. S. Quatrano.

4 recordsLinked to original sources

Localization of Actin mRNA during the Establishment of Cell Polarity and Early Cell Divisions in Fucus Embryos.

Localization of mRNA is a well-described mechanism to account for the asymmetric distribution of proteins in polarized somatic cells and embryos of animals. In zygotes of the brown alga Fucus, F-actin is localized at the site of polar growth and accumulates at the cell plates of the first two divisions of the embryo. We used a nonradioactive, whole-mount in situ hybridization protocol to show the pattern of actin mRNA localization. Until the first cell division, the pattern of actin mRNA localization is identical to that of total poly(A)+ RNA, that is, a symmetrical distribution in the zygote followed by an actin-dependent accumulation at the thallus pole at the time of polar axis fixation. At the end of the first division, actin mRNA specifically is redistributed from the thallus pole to the cell plates of the first two divisions in the rhizoid. This specific pattern of localization in the zygote and embryo involves the redistribution of previously synthesized actin mRNA. The initial asymmetry of actin mRNA at the thallus pole of the zygote requires polar axis fixation and microfilaments but not microtubules, cell division, or polar growth. However, redistribution of actin mRNA from the thallus pole to the first cell plate is insensitive to cytoskeletal inhibitors but is dependent on cell plate formation. The F-actin that accumulates at the rhizoid tip is not accompanied by the localization of actin mRNA. However, maintenance of an accumulation of actin protein at the cell plates of the rhizoid could be explained, at least partially, by a mechanism involving localization of actin mRNA at these sites. The pattern and requirements for actin mRNA localization in the Fucus embryo may be relevant to polarization of the embryo and asymmetric cell divisions in higher plants as well as in other tip-growing plant cells.

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Molecular Responses to Abscisic Acid and Stress Are Conserved between Moss and Cereals.

Promoter elements from the wheat Em gene have been characterized. These elements are inducible by abscisic acid (ABA) and by osmotic stress. In this study, we demonstrated that the same promoter elements function in a distantly related plant species, the moss Physcomitrella patens. Transient and stable expression of the [beta]-glucuronidase reporter gene was used to determine that the heterologous wheat promoter also responds to osmotic stress and ABA in moss. Mutational analysis of the promoter indicated that the mechanism of gene regulation is conserved in both species. Gel retardation and DNase I footprint analyses were conducted to characterize further the interaction of moss transcription factors with the Em promoter. In addition, the synthesis of stress-related polypeptides in moss was observed. The evolutionary significance of these data and the potential for studying the entire ABA perception-response pathway in moss are discussed.

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Actin Localization during Fucus Embryogenesis.

Embryogenesis in the Fucales serves as a model system for studying the acquisition of cellular and developmental polarity. Fertilized eggs bear no asymmetry, yet within 16 hours, a developmental axis is formed and the unicellular zygote germinates in accordance with this axis. Microfilaments (actin) play a crucial role in establishing the axis as evidenced by the inhibitory effects of cytochalasins on axis fixation. The cellular content of actin was determined by immunoblot, whereas the localization of F-actin was investigated using the fluorescent probe rhodamine phalloidin. Three isoforms of actin were detected in constant amounts at all developmental stages. Actin networks were found to be distributed uniformly in eggs and zygotes through the period of early zygote development when the polar axis was formed. However, as the polar axis became irreversibly fixed in space, actin was localized at the presumptive germination site by a cytochalasin-sensitive process. This correlation supports the proposal that actin networks play a critical role in axis fixation, and is consistent with our hypothesis that this process involves stabilization of membrane components by transmembrane bridges from the cell wall to the microfilament cytoskeleton.

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