Unreplicated Chromosomes Research Articles

Large palindromes in the lambda phage genome are preserved in a rec+ host by inhibiting lambda DNA replication.

A large palindrome carried by phage lambda has been shown to prevent growth of the phage on a rec+ strain of Escherichia coli. The phage do form plaques on recBC sbcB strains, but the palindrome is not stable--deletions that either destroy the palindrome or diminish its size overgrow the original engineered palindrome-containing phage. We have prepared stocks of lambda carrying a palindrome that is 2 X 4200 base pairs long. These phage stocks are produced by induction of a lysogen in which the two halves of the palindrome are stored at opposite ends of the prophage and are of sufficient titer (10(9) phage per ml) to enable one-step growth experiments with replication-blocked phage. We find that the large palindrome as well as a lesser palindrome of 2 X 265 base pairs are recovered intact among particles carrying unreplicated chromosomes following such an infection of a rec+ host. We propose that DNA replication drives the extrusion of palindromic sequences in vivo, forming secondary structures that are substrates for the recBC and sbcB gene products.

A model for the recA-dependent repair of excision gaps in UV-irradiated Escherichia coli

We have tested and supported the hypothesis that, in UV-irradiated Escherichia coli, recA-dependent nucleotide excision repair only functions in the replicated portion of the chromosome (i.e., where sister duplexes exist). Using a dnaA(Ts) mutation to align the chromosomes (i.e., all rounds of DNA replication were completed, and new rounds could not be initiated), we studied the genetic control of excision repair (measured as the repair of excision gaps in DNA) in cells with unreplicated chromosomes, and also in cells with partially replicated chromosomes. The excision repair that occurred in cells with unreplicated chromosomes was recA independent, but the excision repair that occurred in cells with partially replicated chromosomes was partially recA dependent. We found no evidence of interchromosomal recombination in recA-dependent excision repair. The majority of this recA-dependent excision repair was recF dependent, and a small portion was recB dependent. The recF and recB genes are suggested to function in excision repair in a manner similar to their function in postreplication repair, i.e., in the replicated portion of a chromosome, the RecF pathway repairs gaps, and the RecB pathway repairs the DNA double-strand breaks that arise at unrepaired gaps.

Cytological studies on degenerative nuclei at pollen development ofCarex ciliato-marginata

Chromosomes in degenerative and functional nuclei ofCarex ciliato-marginata Nakai were investigated during meiotic and primary pollen nuclear division. The nuclear DNA content of these nuclei was also measured using Feulgen microspectrophotometry. At metaphase of the primary pollen nuclear division, the chromosomes of degenerative nuclei were the same length as those of the functional nucleus, but only half their width. The functional nucleus divided into two, each of which moved to a pole, but the degenerative nuclei did not divide. The nuclear DNA content of the degenerative nucleus was half that of the functional nucleus and equal to that of one of the tetrads of a meiotic division. It is concluded that DNA replication was carried out in only one nucleus of the tetrad and that the other three nuclei were composed of unreplicated chromosomes at metaphase of the primary pollen nuclear division.

DNA synthesis at the site of a Red-mediated exchange in phage lambda.

In phage lambda, progeny particles bearing unreplicated chromosomes are recombinant by action of lambda's Red system only near the right end of the chromosome. These recombinants are frequently heterozygous (heteroduplex) for markers located there. In replication-blocked crosses involving two heavy-labeled parents we find that particles in the solitary peak, containing progeny with fully conserved DNA, vary in density. Those on the heavy side of this peak are more apt to be heterozygous than are those on the light side. The data fit a model in which a double chain cut at cos, lambda's packaging origin, is followed by partial exonucleolytic degradation of lambda's r chain from the right end leftward. The exposed l chain, which thereby constitutes a 3' overhang, invades an intact, circular homologue after itself suffering some degradation. Completion of the recombinant chromosome sometimes involves DNA synthesis primed by the invading chain.

The formation and recovery of two-break chromosome rearrangements from irradiated spermatozoa of Drosophila melanogaster

Chromosome and chromatid-type rearrangements can be induced by exposure of spermatozoa of Drosophila to ionising radiation. A model, proposed to explain the formation and recovery of compound autosomes, has been extended to account for the induction of centric fragments capped by a duplication of paternal chromosome material. Three basic assumptions have been used; (1) that the sperm nucleus contains a haploid set of unreplicated chromosomes, (2) that the broken chromosomes ends can be joined together before or after replication, and (3) that one of the first two cleavage nuclei may be lost and an adult organism derived from the other. The present paper reports a theoretical application of this combination of assumptions to the general case of the formation and recovery of two-break rearrangements. This present papers led to an eludidation of the relation between repeats, compounds, fragments, amd deficiencies on the on hand and inversions and translocations on the other hand. Dicentric chromosomes and segmental aneuploidy can be simply explained. A selective screen is formed by the segregation of chromatid rearrangements and the aneuploidy tolerance, Thus there is an alternative way of explaining observations which might indicate preferential breakage or joining.

The distribution of crossovers along unreplicated lambda bacteriophage chromosomes.

The distribution of crossovers along unreplicated chromosomes of bacteriophage lambda has been examined by determining the density distributions and genotypes of particles in the progenies of crosses of density-labeled by ordinary parents in the presence of genetic blocks to replication. The Red and Rec systems combined produce crossovers primarily near the ends (especially the right end) of the chromosome. Removal of the generalized lambda recombination functions by red and gam mutations results in loss of these terminal crossovers; coupled with this loss is a disappearance of the differential dependence of recombination frequencies in terminal and central intervals on DNA synthesis. Removal of the bacterial system by a recA mutation results in severe depression of crossing over among unreplicated phage, with the few recombinants produced by the lambda system occurring near the right end.

A role for recBC nuclease in the distribution of crossovers along unreplicated chromosomes of phage lambda.

The gam gene of phage λ plays a role in the generation of recombinants in terminal intervals among particles bearing unreplicated chromosomes. Its activity in this regard can be entirely accounted for by the reported ability of the wild type gam product to inactivate the product of the recBC genes of the host cell.

LATERAL SUBUNITS IN C-BANDS OF UNREPLICATED CHROMOSOMES OF DISSOSTEIRA CAROLINA (L.) (ORTHOPTERA:ACRIDIDAE)

The C-banding pattern is described for the chromosomes of the grasshopper Dissosteira Carolina (2n♂ = 24, ♀ = 23; XX-XO). Each of the 24 acrocentric chromosomes has a prominent C-band at the centromeric region and most chromosomes have a less prominent C-band at the distal telomeric region. Usually, one or more pairs of chromosomes has a C-band in the short arm. The C-band at the centromeric region appears to be composed of two lateral subunits in unreplicated meiotic chromosomes at anaphase II and telophase II in a number of secondary spermatocytes and in unreplicated mitotic chromosomes at anaphase in several spermatogonia. The C-band at the distal telomeric region appears similarly divided in unreplicated chromatids at anaphase I and metaphase II in several spermatocytes. The lateral subunits may be an artifact or may reflect the presence of a dual structure at C-banded regions.