Polytene chromosomes of Oxytricha: Biochemical and morphological changes during macronuclear development in a ciliated protozoan

Chromosoma ◽  
1976 ◽  
Vol 54 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Brian B. Spear ◽  
Marlene R. Lauth
1973 ◽  
Vol 13 (2) ◽  
pp. 479-509 ◽  
Author(s):  
K. G. MURTI

This report describes an ultrastructural investigation of macronuclear development following conjugation in Stylonychia mytilus (a spirotrichous ciliate) and Tetrahymena pyriformis (a holotrichous ciliate). In S. mytilus, polytene chromosomes are formed in the young macronucleus (macronuclear Anlage). They are subsequently broken between the bands by ‘membranous’ partitions; the assembly of the membranes appears to be concomitant with the formation of the polytene chromosomes. The membranes in the Anlage appear to originate from fibrous material seen in the early Anlage. This fibrous material in the earlier stages is seen concentrated at several points along the border of the inner nuclear membrane. In the later stages it is seen in the interior of the Anlage, outlining the developing polytene chromosomes. As the chromosomes reach the maximum degree of polyteny, the fibrous material condenses to acquire a membranous appearance and extends into the interband regions. The Anlage throughout this period shows a progressive increase in size. Subsequently, the membranes enclose individually each band plus portions of the 2 adjacent interband regions of the polytene chromosomes to form a large number of vesicles. After vesicle formation the Anlage shrinks, and the chromatin inside the vesicles shows degradative changes. Finally, the vesicles disappear, the membrane degradation products appear at the nuclear membrane, and the Anlage now contains nucleoli. The Anlage increases its DNA content by multiple rounds of replication to become a mature macronucleus. The ultra-structural changes observed in the Anlage support the idea of genetic diminution (i.e. extensive DNA synthesis, elimination of many DNA nucleotide sequences, and amplification of the remaining DNA nucleotide sequences in a second period of DNA synthesis) proposed earlier on the basis of cytochemical, biochemical, and limited electron-microscope studies. In T. pyriformis, the macronuclear development differs substantially from that of Stylonychia. Features such as the formation and degradation of polytene chromosomes are absent in the macronuclear development of Tetrahymena; the young macronucleus in this cell becomes a mature macronucleus by progressive increment in size and chromatin content with no apparent genetic diminution. These observations agree with cytochemical studies on the macronuclear development of Tetrahymena.


1970 ◽  
Vol 47 (2) ◽  
pp. 395-407 ◽  
Author(s):  
John A. Kloetzel

The development of the macronucleus following conjugation in the hypotrichous ciliates Euplotes and Stylonychia has been examined with the electron microscope. Banded polytene chromosomes can be seen in thin sections of the macronuclear anlagen during the early periods of exconjugant development. As the chromosomes reach their maximum state of polyteny, sheets of fibrous material appear between the chromosomes and transect the chromosomes in the interband regions. Individual bands of the polytene chromosomes thus appear to be isolated in separate compartments. Subsequently, during the stage when the bulk of the polytenic DNA is degraded (1), these compartments swell, resulting in a nucleus packed with thousands of separate spherical chambers. Individual chromosomes are no longer discernible. The anlagen retain this compartmentalized condition for several hours, at the end of which time aggregates of dense material form within many of the compartments. The partitioning layers disperse shortly before replication bands appear within the elongating anlagen, initiating the second period of DNA synthesis characteristic of macronuclear development in these hypotrichs. The evidence presented here suggests that the "chromatin granules" seen in the mature vegetative macronucleus represent the material of single bands of the polytene chromosomes seen during the earlier stages of macronuclear development. The possibility is also discussed that the degradation of DNA in the polytene chromosomes may be genetically selective, which would result in a somatic macronucleus with a different genetic constitution than that of the micronucleus from which it was derived.


Cell ◽  
1984 ◽  
Vol 36 (4) ◽  
pp. 1045-1055 ◽  
Author(s):  
Lawrence A. Klobutcher ◽  
Carolyn L. Jahn ◽  
David M. Prescott

1995 ◽  
Vol 15 (12) ◽  
pp. 6488-6495 ◽  
Author(s):  
J S Frels ◽  
C L Jahn

Macronuclear development in Euplotes crassus begins with polytenization of micronuclear chromosomes and is accompanied by highly precise excision of DNA sequences known as internal eliminated sequences and transposon-like elements (Tecs). Quantitation of radiolabeled-precursor incorporation into DNA indicates that DNA synthesis during formation of polytene chromosomes is not continuous and occurs during two distinct periods. We demonstrate that the timing of Tec excision coincides with these replication periods and that excision can occur during both periods even at a single locus. We also show that Tec and internal eliminated sequence excisions are coincident in the second replication period, thus providing further evidence for similarity in their excision mechanism. Inhibition of DNA synthesis with hydroxyurea diminishes Tec element excision, indicating that replication is an important aspect of the excision process.


2004 ◽  
Vol 3 (2) ◽  
pp. 288-301 ◽  
Author(s):  
Jeffrey S. Fillingham ◽  
Ronald E. Pearlman

ABSTRACT Extensive programmed DNA rearrangements occur during the development of the somatic macronucleus from the germ line micronucleus in the sexual cycle of the ciliated protozoan Tetrahymena thermophila. Using an in vivo processing assay, we analyzed the role of micronucleus-limited DNA during the programmed deletion of mse2.9, an internal eliminated sequence (IES). We identified a 200-bp region within mse2.9 that contains an important cis-acting element which is required for the targeting of efficient programmed deletion. Our results, obtained with a series of mse2.9-based chimeric IESs, led us to suggest that the cis-acting elements in both micronucleus-limited and macronucleus-retained flanking DNAs stimulate programmed deletion to different degrees depending on the particular eliminated sequence. The mse2.9 IES is situated within the second intron of the micronuclear locus of the ARP1 gene. We show that the expression of ARP1 is not essential for the growth of Tetrahymena. Our results also suggest that mse2.9 is not subject to epigenetic regulation of DNA deletion, placing possible constraints on the scan RNA model of IES excision.


1985 ◽  
Vol 101 (1) ◽  
pp. 79-84 ◽  
Author(s):  
M Roth ◽  
M Lin ◽  
D M Prescott

After conjugation in hypotrichous ciliates, a new macronucleus is produced from a copy of the micronucleus. This transformation involves large-scale reorganization of DNA, with conversion of the chromosomal micronuclear genome into short, gene-sized DNA molecules in the macronucleus. To study directly the changes that occur during this process, we have developed techniques for synchronous mating of large populations of the hypotrichous ciliate Euplotes crassus. Electron microscope studies show that the micronuclear chromosomes are polytenized during the first 20 h of macronuclear development. The polytene chromosomes lack the band-interband organization observed in other hypotrichs and in the Diptera. Polytenization is followed by transectioning of the chromosomes. We isolated DNA at various times of macronuclear development and found that the average molecular weight of the DNA decreases at the time of chromosome transectioning. In addition, we have shown that a small size group of macronuclear DNA molecules (450-550 base pairs) is excised from the chromosomal DNA approximately 10 h later in macronuclear development.


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