Alliegro Mark C.

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Alliegro
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Mark C.
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  • Article
    The karyomastigont as an evolutionary seme
    (University of Chicago Press, 2012-12) Chapman, Michael ; Alliegro, Mark C.
    The problem of eukaryogenesis—the evolutionary mechanism whereby eukaryotic cells evolved from prokaryotes—remains one of the great unsolved mysteries of cell biology, possibly due to the reductionist tendency of most scientists to work only within their subdisciplines. Communication between biologists who conduct research on the nucleus and those working on the cytoskeleton or endomembrane system are sometimes wanting, and yet, all of these quintessentially eukaryotic elements of the cell are interdependent, and are physically associated in many protists as the karyomastigont organellar system: nucleus, one or more basal bodies and flagella, nuclear connector, and Golgi apparatus. Here we suggest a more holistic view of the karyomastigont as not simply an organellar system, but an evolutionary seme, the archaic state of the eukaryotic cell. We also present a scheme whereby the karyomastigont may have dissociated, giving rise in more derived cells to one or more free nuclei and discrete flagellar apparati (akaryomastigonts).
  • Preprint
    Rediscovery of the nucleolinus, a dynamic RNA-rich organelle associated with the nucleolus, spindle, and centrosomes
    ( 2010-06) Alliegro, Mary Anne ; Henry, Jonathan J. ; Alliegro, Mark C.
    The nucleolinus is an RNA-rich compartment, closely apposed to or embedded within the nucleolus. Discovered over 150 years ago, fewer than two dozen articles have been published on the nucleolinus, probably because complex histochemical stains are required for its visualization in the great majority of cells. The nucleolinus has been reported in invertebrate oocytes, mammalian and amphibian epithelial cells, neurons, and several transformed cell lines. A prominent nucleolinus, clearly visible with transmitted light microscopes at 10x magnification, is present in each oocyte of the surf clam, Spisula solidissima. We observed a consistent relationship between the nucleolinus and the developing meiotic apparatus following Spisula oocyte activation. Through sonication and sucrose gradient fractionation of purified oocyte nuclei, we isolated nucleolini, extracted their RNA, and prepared an in situ riboprobe (NLi-1) that is associated specifically with the nucleolinus, confirming its unique composition. Other in situ observations revealed a NLi-1 and nucleolinar association with the developing spindle and centrosomes. Laser microsurgery that targeted the nucleolinus resulted in failed meiotic cell division in parthenogenetically activated oocytes and failed mitosis in fertilized oocytes. Although the nucleolinus may be a forgotten organelle, its demonstrated role in spindle formation suggests it deserves renewed attention.
  • Preprint
    The centrosome and spindle as a ribonucleoprotein complex
    ( 2011-01-03) Alliegro, Mark C.
    The presence of nucleic acids in centrosomes and the spindle have been proposed, observed, and reported since the 1950s. Why did the subject remain, perhaps even until today, such a controversial issue? The explanation is manifold, and includes legitimate concern over contamination from other cellular compartments in biochemical preparations. With a typically high background of cytoplasmic ribosomes, even microscopic images of stained intact cells could be difficult to interpret. Also, evidence for RNA and DNA in centrosomes accumulated for approximately 40 years but was interspersed with contradictory studies, primarily regarding the presence of DNA (reviewed in Johnson and Rosenbaum, 1991; Marshall and Rosenbaum, 2000). Perhaps less tangible but still a likely cause for lingering controversy is that the presence of nucleic acids in the spindle or centrosomes will require us to look differently at these structures from a functional, and more to the point, evolutionary standpoint.
  • Preprint
    Composition and dynamics of the nucleolinus, a link between the nucleolus and cell division apparatus in surf clam (Spisula) oocytes
    ( 2011-12-26) Alliegro, Mark C. ; Hartson, Steven ; Alliegro, Mary Anne
    The nucleolinus is a little-known cellular structure, discovered over 150 years ago (1) and thought by some investigators in the late 19th to mid-20th century to function in the formation of the centrosomes or spindle. A role for the nucleolinus in formation of the cell division apparatus has recently been confirmed in oocytes of the surf clam, Spisula solidissima (2). However, we know so little about the composition and dynamics of this compartment, it is difficult to construct mechanistic hypotheses or even to be sure that prior reports were describing analogous structures in the cells of mammals, amphibians, plants, and other organisms where it was observed. Surf clam oocytes are an attractive model to approach this problem because the nucleolinus is easily visible by light microscopy, making it accessible by laser microsurgery as well as isolation by common cell fractionation techniques. In this report we analyze the macromolecular composition of isolated Spisula nucleolini and examine the relationship of this structure to the nucleolus and cell division apparatus. Analysis of nucleolinar RNA and protein revealed a set of molecules that overlaps with, but is nevertheless distinct from the nucleolus. The proteins identified were primarily ones involved in nucleic acid metabolism and cell cycle regulation. Monoclonal antibodies generated against isolated nucleolini revealed centrosomal forerunners in the oocyte cytoplasm. Finally, induction of damage to the nucleolinus by laser microsurgery altered the trafficking of α- and γ-tubulin after fertilization. These observations strongly support a role for the nucleolinus in cell division and represent our first clues regarding mechanism.