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(NAS Colloquium) Molecular Kinesis in Cellular Function and Plasticity (2002)
National Academy of Sciences (NAS)

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. "Spatial and temporal control of RNA stability." (NAS Colloquium) Molecular Kinesis in Cellular Function and Plasticity. Washington, DC: The National Academies Press, 2002.

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Colloquium on Molecular Kinesis in Cellular Function and Plasticity

Fig. 1. Time course of maternal transcript degradation in activated, unfertilized eggs. (A) The same Northern blot probed for rpA1 (stable) and string, Hsp83, or nanos (unstable) transcripts. (B) Quantitative analysis of the time course of Hsp83 transcript degradation. The points represent the ratio of Hsp83 transcripts to stable rpA1 transcripts relative to the initial (0-0.5 h) concentration. It can be seen that more than 95% of the Hsp83 transcripts have disappeared by 3.0 to 3.5 h after egg activation. Data from two independent experiments are presented. Half-hour time windows are shown. See ref. 9 for details.

lated region (UTR) of Hsp83 and nanos that, when deleted, result in substantial stabilization of corresponding transgenic transcripts in both unfertilized and fertilized eggs (Fig. 2) (9, 14-16). It was comparison of the stability of the cis element-deleted transcripts in unfertilized versus fertilized eggs that led to the discovery of the zygotic degradation pathway (9): transcripts deleted for an element required for maternal degradation are fully stabilized in unfertilized eggs (Fig. 2 A) but are destabilized starting 2 h after fertilization in developing embryos (Fig. 2 B). Thus, there must be additional cis-acting elements that are still present in these transcripts and that mediate zygotic degradation.

Spatial Control of Maternal RNA Stability in Drosophila Although unstable maternal transcripts such as string and Hsp70 are eliminated throughout the egg or early embryo (Fig. 3 A and B), unstable transcripts such as Pgc, Hsp83, and nanos are eliminated from the bulk cytoplasm of the egg or embryo but remain stable at the posterior (Fig. 3 C-F) (9). Uniform instability is the default state for the unstable classes of transcripts: if the Hsp83 3′ UTR is replaced with a 3′ UTR from a uniformly degraded transcript (e.g., Hsp70) or if a cis-acting "protection" element is deleted (see below), then the resulting transgenic Hsp83 transcripts are degraded throughout the embryo (see Fig. 5

Fig. 2. Removal of a maternal Hsp83 degradation element stabilizes transgenic transcripts in unfertilized (A) but not fertilized (B) eggs. Northern blots are shown that were simultaneously probed for (i) endogenous Hsp83 transcripts; (ii) transgenic reporter transcripts carrying the Hsp83 5′ UTR, the first 111 codons of the Hsp83 ORF, an Escherichia coli ß-galactosidase RNA tag, and the Hsp83 3' UTR deleted for a 97-nt element referred to as the Hsp83 degradation element (HDE) (for a detailed description of this transgene see ref. 9); (iii) endogenous rpA1 transcripts. On both blots it can be seen that endogenous Hsp83 transcripts are unstable and endogenous rpA1 transcripts are stable. However, although transgenic ∆HDE transcripts are stable in unfertilized eggs (1A), they are degraded commencing 2 h after fertilization in developing embryos (B). Half-hour time windows after egg activation or fertilization are shown. See ref. 9 for details.

A and B) (9). This experiment proves that the endogenous Hsp83, nanos, and Pgc transcripts that remain at the posterior of the egg and early embryo are protected from degradation in that region of the cytoplasm (i.e., the degradation machinery is

Fig. 3. Certain classes of maternal transcripts are degraded throughout the cytoplasm of activated, unfertilized eggs whereas others are protected from degradation in the posterior polar plasm, string transcripts are initially present throughout the egg (A) and are subsequently degraded (B). In contrast, whereas Hsp83 (C) and nanos (E) transcripts are initially present in both the posterior polar plasm and the presumptive somatic region (C and E), degradation is limited to the somatic region whereas transcripts are protected from degradation in the posterior polar plasm (D and F). (A, C, and E) One to 2 h after egg activation; (B, D, and F) 3-4 h after egg activation. Whole-mount RNA in situ hybridizations are shown, with anterior to the left and dorsal toward the top of the page. See ref. 9 for details.

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Front Matter (R1-R4)
Introduction: Molecular kinesis in cellular function and plasticity (1-2)
Kinesin molecular motors: Transport pathways, receptors, and human disease (3-7)
All kinesin superfamily protein, KIF, genes in mouse and human (8-15)
Assembly and transport of a premessenger RNP particle (16-21)
Ribonucleoprotein infrastructure regulating the flow of genetic information between the genome and the proteome (22-28)
Spatial and temporal control of RNA stability (29-32)
Molecular mechanisms of translation initiation in eukaryotes (33-40)
The target of rapamycin (TOR) proteins (41-48)
The physiological significiance of ß-actin mRNA localization in determining cell polarity and directional motility (49-54)
Sorting and directed transport of membrane proteins during development of hippocampal neurons in culture (55-61)
Molecular organization of the postsynaptic specialization (62-65)
A cellular mechanism for targeting newly synthesized mRNAs to synaptic sites on dendrites (66-72)
Think globally, translate locally: What mitotic spindles and neuronal synapses have in common (73-75)
Vasopressin mRNA localization in nerve cells: Characterization of cis-acting elements and trans-acting factors (76-83)
Local translation of classes of mRNAs that are targeted to neuronal dendrites (84-89)
Cytoskeletal microdifferentiation: A mechanism for organizing morphological plasticity in dendrites (90-96)
Tracking the estrogen receptor in neurons: Implications for estrogen-induced synapse formation (97-104)
Synaptic regulation of protein syntesis and the fragile X protein (105-110)
Proceedings program (111-112)