Association of 7 SL RNA and an SRP-like particle with polysomes and endoplasmic reticulum in the developing sea urchin embryo.
We have identified the sea urchin cognate of the mammalian signal recognition particle (SRP). This particle contains the diagnostic 7 SL small RNA, sediments at a similar velocity to that reported for the mammalian particle, and is found associated with the ER and polysomes. We have examined its subcellular localization during embryogenesis in order to determine whether it could serve in a translational regulatory capacity for a subset of the stored maternal mRNAs. In these studies the 7 SL RNA was used as a marker for the particle, since we determined that the 7 SL RNA exists exclusively within the SRP-like particle at all developmental stages. The relative distribution of the SRP among cytoplasmic structures changes dramatically during development. This represents an actual change in subcellular localization because the 7 SL RNA level remains nearly constant per embryo until the pluteus stage, when it increases slightly. In eggs, the SRP exists almost entirely free in the cytoplasm as an 11 S particle. Very soon after fertilization and throughout development there is an increase in the association of the particle with rapidly sedimenting structures, until by the pluteus stage greater than 90% of the SRP exists in a bound state. The nature of the associations is complex, and the bound structures include, at least in part, ribosomes, polysomes, and microsomes. The SRP is associated with microsomal membranes in gastrula (36 hr) but not in blastula (12 hr) or earlier embryos. Using the criteria of sensitivity to Triton X-100, we determined that 16% of the SRP in a 10,000g cytoplasmic fraction was bound to membranes in a microsomal (endoplasmic reticulum)-containing fraction in the gastrula. In contrast, less than 1% was membrane associated in the blastula. The SRP was also found in a ribosome-polysome fraction in 12-, 36-, and 48-hr embryos, but not in eggs. Finally, a small but significant portion of the SRP was found associated with monosomes in cleavage stage embryos. The possible role the SRP could play in the elongation arrest of stored maternal messages for secreted proteins is discussed.
LeBlanc
J M
JM
Molecular Biology and Biochemistry Department, Wesleyan University, Middletown, Connecticut 06457.
Infante
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AA
D000818:Animals; D015152:Blotting, Northern; D002451:Cell Compartmentation; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D004492:Edetic Acid / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000502:physiology*; D005306:Fertilization; D005775:Gastrula / Q000502:physiology; D066298:In Vitro Techniques; D008861:Microsomes / Q000378:metabolism; D011132:Polyribosomes / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000502:physiology*; D012617:Sea Urchins / Q000196:embryology* / Q000502:physiology; D018271:Signal Recognition Particle
D012261:Ribonucleoproteins; D018271:Signal Recognition Particle; D012313:RNA; D004492:Edetic Acid