Hematopoietic stem cell lodgment in the adult bone marrow stem cell niche.

Treatment of malignant blood disorders, such as leukemia, that can provide a better chance of long-term remission involves myeloablation followed by transplantation of matched donor hematopoietic stem cells (HSCs). For successful engraftment and re-establishment of hematopoiesis to occur in the recipient, the transplanted HSCs must first migrate from the blood circulation to the bone marrow (BM), a process known as homing, then localize and anchor in suitable microenvironments within the BM, a process known as lodgment. After lodgment, the specific fate of the transplanted HSCs is determined through complex, bidirectional interactions with various stromal cell components in the niche. Ultimately, these interactions dictate the clinical outcome of the transplantation. Through the use of transgenic mouse models, considerable evidence has been accumulated in an attempt to unveil the possible underlying mechanisms that govern these processes. Here, we will emphasize the major factors that are involved in the regulation of lodgment of transplanted HSCs. Specifically, we will first introduce early observations on the spatial distribution of hematopoietic progenitors within the BM, then we will discuss the soluble factors, chemokines, cell-cell interactions, and cell-matrix interactions that have been studied and known to influence the site of HSC lodgment within the BM following transplantation.

Transposon mutagenesis and tagging of fluorescent Pseudomonas: Antimycotic production is necessary for control of Dutch elm disease.

Antimycotic-producing strains of Pseudomonas syringae are being tested as Dutch elm disease control agents. We examined the role of antimycotic production in disease control. Transposon Tn903 was used to mutagenize the antimycotic-producing strain MSU174. Eighty-one mutants that did not inhibit fungal growth were identified among 15,000 Tn903-containing derivatives. Linkages between Tn903 insertions and defects in antimycotic metabolism were established. Three Tn903-containing strains (two antimycotic producers and one nonproducer) were individually introduced into American elm seedlings. The seedlings were subsequently challenged with Ceratocystis ulmi, the causal agent of Dutch elm disease. Protection of the elms was observed with the two antimycotic-producing strains but not with the nonproducing strain. The introduced strains could be readily recovered from the seedlings after two growing seasons. They were unequivocally identified by the Tn903 insertions they contain.

A vehicle for the introduction of transposons into plant-associated pseudomonads.

A recombinant plasmid with wide-host-range transfer functions, narrow-host-range replication functions, and carrying a kanamycin-resistant transposon transferred kanamycin resistance to a number of plant-associated pseudomonads. Southern hybridization studies suggest that only a small portion of the plasmid, coinciding with the location of the transposon, is present in the kanamycin-resistant Pseudomonas derivatives. The plasmid sequences appear to be inserted at a number of different sites in the recipient genome. This plasmid can thus be used as a vehicle for the introduction of transposons into some plant-associated pseudomonads and should be useful in both genetic and ecological studies of these bacteria.

Isolated clusters of paired tandemly repeated sequences in the Xenopus laevis genome.

There exist in the Xenopus laevis genome clusters of tandemly repeated DNA sequences, consisting of two types of 393-base-pair repeating unit. Each such cluster contains several units of one of these paired tandem repeats (PTR-1), followed by several units of the other repeat (PTR-2). The number of repeats of each type is variable from cluster to cluster and averages about seven of each type per cluster. Every cluster has ca. 1,000 base pairs of common left flanking sequence (adjacent to the PTR-1 repeats) and 1,000 base pairs of common right flanking sequence (adjacent to the PTR-2 repeats). Beyond these common flanks, the DNA sequences are different in the eight cloned genomic fragments we have studied. Thus, the hundreds of PTR clusters in the genome are dispersed at apparently unrelated sites. Nucleotide sequences of representative PTR-1 and PTR-2 repeats are 64% homologous. These sequences do not reveal an obvious function. However, the related species X. mulleri and X. borealis have sequences homologous to PTR-1 and PTR-2, which show the same repeat lengths and genomic organization. This evolutionary conservation suggests positive selection for the clusters. Maintenance of these sequences at dispersed sites imposes constraints on possible mechanisms of concerted evolution.

Tandemly repeated DNA sequences from Xenopus laevis. II. Dispersed clusters of a 388 base-pair repeating unit.

A repetitive DNA sequence family from Xenopus laevis that has an unusual genomic organization has been identified. It has been shown by blot-hybridization that this sequence occurs in clusters containing variable numbers of the 388 base-pair repeating unit. There are approximately 500 such clusters in the genome, and each cluster has common flanking sequences. The number of tandem 388 base-pair repeats per cluster ranges from one to at least 15, with a mean of seven. Homologous sequences were found in two related species, Xenopus borealis and Xenopus mulleri, where the size of the repeating units and their genomic arrangement are very similar to those in X. laevis. The complete nucleotide sequence of a cloned representative 388 base-pair repeating unit showed no short internal repeats and no long reading frames. By blot-hybridization, no evidence of transcripts of this sequence was found in total RNA from X. laevis liver, embryos or oocytes.

Tandemly repeated DNA sequences from Xenopus laevis. I. Studies on sequence organization and variation in satellite 1 DNA (741 base-pair repeat).

A family of tandemly repeated sequences, having a basic repeating unit of 741 base-pairs, has been identified in Xenopus laevis DNA and designated satellite 1. Apart from its rather long repeat unit, the characteristics of this DNA appear to be quite similar to those of complex satellite DNAs from other organisms. The nucleotide sequence of a cloned repeat unit shows no evidence of simpler internal repeats, and there is no obvious suggestion of reasonable RNA- or protein-coding regions. Transcripts homologous to this DNA could not be demonstrated in liver, embryo or oocyte RNAs. By blot-hybridization, satellite 1 has been shown to exist in the genome chiefly as tandem repeats of the 741 base-pair sequence. However, there are a number of repeats that differ from the normal sequence (as judged by loss or gain of restriction sites) and some that differ in length. Similar variants are often, but not always, clustered. Characterization of genomic clones of this satellite has confirmed the tandem organization and clustering of variants. The nature of some variants has been elucidated in more detail. Some regions of the basic repeat seem to be more prone to variation than are others.