PCB congener distributions in muscle, liver and gonad of Fundulus heteroclitus from the lower Hudson River Estuary and Newark Bay.

Gradients in sediment polychlorinated biphenyl (PCB) concentrations and PCB congener profiles exist along the Hudson River (NY, USA). We evaluated site and tissue differences in PCB concentration and congener profiles in resident mummichog (Fundulus heteroclitus) collected from PCB-contaminated sites in the lower Hudson River and the New York/New Jersey Harbor Estuary. Fish were collected from three PCB-contaminated sites Piermont Marsh (P), Iona Marsh (I), and Newark Bay (NB), and from two reference sites (Flax Pond, NY; Succotash Salt Marsh, RI). Congener profiles were statistically analyzed using principal component analysis (PCA) and general linear model (GLM) profile analysis. Contaminated fish had PCB tissue concentrations approximately 10-fold higher than those of reference fish. There were no site differences in PCB body burden (all tissues combined) among the contaminated site fish. However, relative PCB concentration did differ between organs: NB fish (gonad=liver>muscle); I and P fish (gonad>liver>muscle). In contrast to PCB content, PCB congener profiles did show site differences; NB mummichog being depleted in the less chlorinated congeners relative to I and P fish, likely reflecting different PCB sources to these populations. Within a site, however, PCB congener patterns were similar between liver, gonad and muscle. In conclusion, PCA and GLM analyses gave complementary results, both analyses indicating differences in site, but not tissue, distributions of PCB congeners. This study also demonstrates that unlike congener profiles, total PCB content does differ dramatically amongst tissues and further, that PCB differences among tissues (gonad vs. liver vs. muscle) can vary with site.

Meiotic segregation, synapsis, and recombination checkpoint functions require physical interaction between the chromosomal proteins Red1p and Hop1p.

In yeast, HOP1 and RED1 are required during meiosis for proper chromosome segregation and the consequent formation of viable spores. Mutations in either HOP1 or RED1 create unique as well as overlapping phenotypes, indicating that the two proteins act alone as well as in concert with each other. To understand which meiotic processes specifically require Red1p-Hop1p hetero-oligomers, a novel genetic screen was used to identify a single-point mutation of RED1, red1-K348E, that separates Hop1p binding from Red1p homo-oligomerization. The Red1-K348E protein is stable, phosphorylated in a manner equivalent to Red1p, and undergoes efficient homo-oligomerization; however, its ability to interact with Hop1p both by two-hybrid and coimmunoprecipitation assays is greatly reduced. Overexpression of HOP1 specifically suppresses red1-K348E, supporting the idea that the only defect in the protein is a reduced affinity for Hop1p. red1-K348E mutants exhibit reduced levels of crossing over and spore viability and fail to undergo chromosome synapsis, thereby implicating a role for Red1p-Hop1p hetero-oligomers in these processes. Furthermore, red1-K348E suppresses the sae2/com1 defects in meiotic progression and sporulation, indicating a previously unknown role for HOP1 in the meiotic recombination checkpoint.

Conserved properties between functionally distinct MutS homologs in yeast.

In the yeast Saccharomyces cerevisiae there are five nuclear MutS homologs that act in two distinct processes. MSH2, 3, and 6 function in mismatch repair in both vegetative and meiotic cells, whereas MSH4 and MSH5 act specifically to facilitate crossovers between homologs during meiosis. Coimmunoprecipitation as well as two-hybrid experiments indicate that the Msh4 and Msh5 proteins form a hetero-oligomeric structure similar to what is observed for the Msh proteins involved in mismatch repair. Mutation of conserved amino acids in the NTP binding and putative helix-turn-helix domains of Msh5p abolish function but are still capable of interaction with Msh4p, suggesting that NTP binding plays a role downstream of hetero-oligomer formation. No hetero-oligomers are observed between the mismatch repair MutS proteins (Msh2p and Msh6p) and either Msh4p or Msh5p. These results indicate that one level of functional specificity between the mismatch repair and meiotic crossover MutS homologs in yeast is provided by the ability to form distinct hetero-oligomers.