TABLE 3.
Transcripts reduced or enhanced more than twofold between J2670 (whiI+) and J2676 (ΔwhiI) at 48 h and/or 72 h of growth
| Open reading framea,f | Genea | Producta | Time of growth (h)b | Normalized intensity ratioc
|
Transcriptional ratiod
|
||||
|---|---|---|---|---|---|---|---|---|---|
| J2670 (whiI+) | J2671 (D27A) | J2673 (D69E) | J2676 (ΔwhiI) | J2670/ J2676 | J2670/ J2671 | ||||
| Gene expression reduced only in whiI null mutantd | |||||||||
| SCO0323* | Unknown | 48 | 4.3 | 4.0 | 6.8 | 0.8 | 5.4 | 1.1 | |
| SCO2718* | rdlA | Secreted protein | 48 | 5.5 | 7.1 | 11.2 | 1.3 | 4.2 | 0.8 |
| SCO0834 | Unknown | 72 | 2.1 | 1.5 | 2.2 | 1.0 | 2.1 | 1.4 | |
| SCO6375* | Secreted protein | 72 | 1.6 | 1.1 | 3.2 | 0.6 | 2.7 | 1.4 | |
| SCO6744 | Unknown | 72 | 1.4 | 1.2 | 1.5 | 0.6 | 2.3 | 1.2 | |
| Gene expression reduced in both whiI null and D27A mutantsd | |||||||||
| SCO4104 | Integral membrane protein | 48 | 2.0 | 1.3 | 1.5 | 1.0 | 2.0 | 1.5 | |
| SCO4393 | Phosphoheptose isomerase | 48 | 3.6 | 2.0 | 2.5 | 1.6 | 2.3 | 1.8 | |
| SCO4761 | groES | 10-kDa chaperonin Cpn10 | 48 | 3.9 | 2.3 | 2.5 | 1.8 | 2.2 | 1.7 |
| SCO5315* | whiE | Polyketide cyclase | 48 | 2.2 | 1.0 | 3.0 | 0.7 | 3.1 | 2.2 |
| SCO6197 | Secreted protein | 48 | 1.2 | 0.6 | 1.1 | 0.6 | 2.0 | 2.0 | |
| SCO0681 | Ferredoxin/ferredoxin-NADP reductase | 72 | 3.7 | 2.4 | 2.2 | 1.5 | 2.5 | 1.5 | |
| SCO0682 | Unknown | 72 | 3.6 | 2.2 | 2.7 | 1.6 | 2.3 | 1.6 | |
| SCO1366* | PadR-like transcriptional regulator | 72 | 7.2 | 3.2 | 3.5 | 2.4 | 3.0 | 2.3 | |
| SCO1367 | ABC transport system ATP-binding protein | 72 | 2.6 | 1.2 | 2.7 | 0.7 | 3.7 | 2.2 | |
| SCO2008* | Branched-chain amino acid binding protein | 72 | 6.4 | 2.4 | 4.4 | 2.1 | 3.0 | 2.7 | |
| SCO2009 | Branched-chain amino acid transport permease | 72 | 1.7 | 1.0 | 1.3 | 0.8 | 2.1 | 1.7 | |
| SCO2162 | Quinolinate synthetase | 72 | 2.5 | 0.7 | 1.8 | 0.8 | 3.1 | 3.6 | |
| SCO3899 | myo-Inositol-1-phosphate synthase | 72 | 3.0 | 1.0 | 1.8 | 1.3 | 2.3 | 3.0 | |
| SCO3900 | Conserved hypothetical protein | 72 | 4.0 | 1.4 | 2.7 | 1.7 | 2.4 | 2.9 | |
| SCO7510 | cypH | Peptidyl-prolyl cis-trans isomerase | 72 | 3.6 | 1.7 | 4.2 | 1.5 | 2.4 | 2.1 |
| SCO7511 | gap-2 | Glyceraldehyde 3-phosphate dehydrogenase | 72 | 3.6 | 1.7 | 4.3 | 1.7 | 2.1 | 2.1 |
| Gene expression reduced in whiI null mutant, with indeterminate response to other whiI allelese | |||||||||
| SCO4172* | Membrane protein | 48 | 1.7 | 0.7 | 0.7 | 0.4 | 4.3 | 2.4 | |
| SCO4174* | Integral membrane protein | 48 | 1.3 | 0.4 | 0.7 | 0.3 | 4.3 | 3.3 | |
| Gene expression enhanced only in whiI null mutantd | |||||||||
| SCO4187 | Membrane protein | 48 | 1.0 | 1.1 | 1.2 | 2.0 | 2.0 | 1.8 | |
| SCO5583 | amtB | Ammonium transporter | 48 | 1.3 | 0.8 | 0.8 | 3.2 | 2.5 | 4.0 |
| SCO5584 | glnK | Nitrogen regulatory protein P-II | 48 | 1.2 | 0.9 | 0.8 | 3.6 | 3.0 | 4.0 |
| SCO3299 | rtcB | Unknown | 72 | 0.6 | 0.5 | 0.6 | 1.3 | 2.2 | 2.6 |
| SCO4002* | Secreted protein | 72 | 1.2 | 2.5 | 2.0 | 4.5 | 3.8 | 1.8 | |
| SCO5189* | Unknown | 72 | 6.3 | 10.2 | 6.9 | 20.5 | 3.3 | 2.0 | |
| SCO5190* | wblC | DNA-binding protein | 72 | 3.8 | 4.6 | 3.7 | 10.9 | 2.9 | 2.4 |
| SCO5191 | Unknown | 72 | 6.9 | 10.0 | 5.5 | 23.5 | 3.4 | 2.4 | |
| SCO7211 | Secreted glycosyl hydrolase | 72 | 0.6 | 1.0 | 1.0 | 1.7 | 2.8 | 1.7 | |
| Gene expression enhanced in both whiI null and D27A mutants | |||||||||
| SCO5249* | Cyclic nucleotide-binding protein | 48 | 2.0 | 2.9 | 2.0 | 4.4 | 2.2 | 1.5 | |
| SCO0140* | MerR family transcriptional regulator | 72 | 0.5 | 1.3 | 0.6 | 1.9 | 3.8 | 1.5 | |
| SCO1648 | arc | AAA ATPase | 72 | 2.5 | 5.2 | 3.0 | 5.0 | 2.0 | 1.0 |
| SCO2922 | Membrane protein | 72 | 1.2 | 3.4 | 0.8 | 2.4 | 2.0 | 0.7 | |
| SCO3123 | prsA2 | Ribose-phosphate pyrophosphokinase | 72 | 1.0 | 1.4 | 1.2 | 2.1 | 2.1 | 1.5 |
| SCO3660 | Integrase/recombinase | 72 | 0.5 | 0.8 | 0.5 | 1.0 | 2.0 | 1.3 | |
| SCO4602 | nuoH2 | NADH dehydrogenase subunit | 72 | 1.0 | 2.2 | 1.2 | 2.6 | 2.6 | 1.2 |
| SCO4605 | nuoK2 | NADH dehydrogenase subunit | 72 | 2.4 | 4.8 | 3.1 | 7.0 | 2.9 | 1.5 |
| SCO5124* | Hypothetical protein | 72 | 2.6 | 6.5 | 2.8 | 6.9 | 2.6 | 1.1 | |
| SCO5125 | Chitinase promoter binding protein 1 | 72 | 1.8 | 3.7 | 2.0 | 3.7 | 2.1 | 1.0 | |
| SCO5269 | Unknown | 72 | 0.9 | 1.9 | 0.9 | 1.8 | 2.0 | 0.9 | |
| SCO5576* | acy | Acylphosphatase | 72 | 1.6 | 4.8 | 2.0 | 4.5 | 2.8 | 0.9 |
| Gene expression enhanced in whiI null mutant, with indeterminate response to other whiI allelese | |||||||||
| SCO1201 | Reductase | 72 | 1.0 | 1.7 | 1.5 | 2.0 | 2.0 | 1.2 | |
The SCO numbers were taken from EMBL database accession no. AL645882. The possible function of each product was either known from previous work or was based on the genome annotation and further information given in the ScoDB database (http://streptomyces.org.uk). Genes in bold are also represented in Streptomyces avermitilis, based on reciprocal hits in BLAST searches.
Earliest culture age showing whiI influence.
The normalization process was performed by the per spot and per chip intensity-dependent (Lowess) normalization method, which performs an intensity-dependent (Lowess) normalization and divides the signal channel (Cy3 channel for cDNA) by the control channel (Cy5 channel for genomic DNA). The values in the table were averaged from all the biological and experimental replicates.
A J2670/J2676 ratio of >2 was the threshold for reduced expression in the whiI null mutant. If the J2670/J2671 ratio was <1.5, then the effect was considered whiI null specific. A J2676/J2670 ratio of >2 was the threshold for enhanced expression in the whiI null mutant, and if the J2676/J2671 ratio was >1.5, then the effect was considered whiI null specific.
For SCO4172 and -4174, expression in J2673 was not consistent with that in J2670 (wild type); for SCO1201, expression in both J2671 and J2673 was intermediate between that in J2670 and J2676.
Genes indicated with asterisks passed all filters during the screen for changes in expression by GeneSpring, whereas the others had differences in at least four of five comparisons (five wild-type replicates, including replicate cultures and experiments, compared with three null mutant individual samples).