Activation of the G2/M-specific gene CLB2 requires multiple cell cycle signals.

In budding yeast (Saccharomyces cerevisiae), the periodic expression of the G2/M-specific gene CLB2 depends on a DNA binding complex that mediates its repression during G1 and activation from the S phase to the exit of mitosis. The switch from low to high expression levels depends on the transcriptional activator Ndd1. We show that the inactivation of the Sin3 histone deacetylase complex bypasses the essential role of Ndd1 in cell cycle progression. Sin3 and its catalytic subunit Rpd3 associate with the CLB2 promoter during the G1 phase of the cell cycle. Both proteins dissociate from the promoter at the onset of the S phase and reassociate during G2 phase. Sin3 removal coincides with a transient increase in histone H4 acetylation followed by the expulsion of at least one nucleosome from the promoter region. Whereas the first step depends on Cdc28/Cln1 activity, Ndd1 function is required for the second step. Since the removal of Sin3 is independent of Ndd1 recruitment and Cdc28/Clb activity it represents a unique regulatory step which is distinct from transcriptional activation.

Physiological responses of bryophytes Thuidium tamariscinum and Hylocomium splendens to increased nitrogen deposition.

BACKGROUND AND AIMS: Increased levels of nitrogen (N) deposition lead to enhanced N contents and reduced productivity of many bryophyte species. This study aimed at elucidating the mechanisms by which enhanced N uptake may cause growth reduction of bryophytes, focusing on the effects of N addition on carbon (C) metabolism of bryophytes. METHODS: Plantlets of Thuidium tamariscinum and Hylocomium splendens were fertilized with NH(4)NO(3) (N load equalling 30 kg ha(-1) year(-1)) for 80 d, including a pulse labelling experiment with (13)CO(2) to dissect the partitioning of carbon in response to N addition. KEY RESULTS: Growth of T. tamariscinum was not affected by N addition, while H. splendens showed a trend towards growth reduction. Total N concentration was significantly increased by N addition in H. splendens, a significant increase in amino acid-N was found in T. tamariscinum only. In both bryophyte species, a reduction in concentration of lipids, the greatest C storage pool, as well as markedly enhanced turnover rates of C storage pools in fertilized plants were observed. CONCLUSIONS: The results suggest that growth reduction of H. splendens under high levels of N deposition may be caused by enhanced synthesis of N-containing organic compounds, most probably of cell wall proteins. Disturbance of cellular C metabolism, as indicated by enhanced C pool turnover, may further contribute to the decline in productivity of H. splendens.

Forkhead-like transcription factors recruit Ndd1 to the chromatin of G2/M-specific promoters.

Many cell-cycle-specific events are supported by stage-specific gene expression. In budding yeast, at least three different nuclear factors seem to cooperate in the periodic activation of G2/M-specific genes. Here we show, by using chromatin immunoprecipitation polymerase chain reaction assays, that a positive regulator, Ndd1, becomes associated with G2/M promoter regions in manner that depends on the stage in cell cycle. Its recruitment depends on a permanent protein-DNA complex consisting of the MADS box protein, Mcm1, and a recently identified partner Fkh2, a forkhead/winged helix related transcription factor. The lethality of Ndd1 depletion is suppressed by fkh2 null mutations, which indicates that Fkh2 may also have a negative regulatory role in the transcription of G2/M-induced RNAs. We conclude that Ndd1-Fkh2 interactions may be the transcriptionally important process targeted by Cdk activity.

Histone deacetylase 1 can repress transcription by binding to Sp1.

The members of the Sp1 transcription factor family can act as both negative and positive regulators of gene expression. Here we show that Sp1 can be a target for histone deacetylase 1 (HDAC1)-mediated transcriptional repression. The histone deacetylase inhibitor trichostatin A activates the chromosomally integrated murine thymidine kinase promoter in an Sp1-dependent manner. Coimmunoprecipitation experiments with Swiss 3T3 fibroblasts and 293 cells demonstrate that Sp1 and HDAC1 can be part of the same complex. The interaction between Sp1 and HDAC1 is direct and requires the carboxy-terminal domain of Sp1. Previously we have shown that the C terminus of Sp1 is necessary for the interaction with the transcription factor E2F1 (J. Karlseder, H. Rotheneder, and E. Wintersberger, Mol. Cell. Biol. 16:1659-1667, 1996). Coexpression of E2F1 interferes with HDAC1 binding to Sp1 and abolishes Sp1-mediated transcriptional repression. Our results indicate that one component of Sp1-dependent gene regulation involves competition between the transcriptional repressor HDAC1 and the transactivating factor E2F1.