Bray, D.

Kapoor, M.

English

Kansas State University

Fungal Genetics Reports Volume 14 Article 3 Catabolite effects on enzymes M. Kapoor D. Bray Follow this and additional works at: https://newprairiepress.org/fgr This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. Recommended Citation Kapoor, M., and D. Bray (1969) "Catabolite effects on enzymes," Fungal Genetics Reports: Vol. 14, Article 3. https://doi.org/10.4148/1941-4765.2028 This Research Note is brought to you for free and open access by New Prairie Press. It has been accepted for inclusion in Fungal Genetics Reports by an authorized administrator of New Prairie Press. For more information, please contact cads@k-state.edu. Catabolite effects on enzymes Abstract Catabolite effects on enzymes This research note is available in Fungal Genetics Reports: https://newprairiepress.org/fgr/vol14/iss1/3 Smith, 6  R The effect of the The recombination-3 gene  described by D. G. Cotcheride (1966 Aurtral.  J.recombination-3 gene on hirtidine-5.Bid.  Sci.  19: 1039) controls the frequency of recombination between pairs ofauxotrophic omination alleles in such a way that  crosser  beoring  the dominant ret-3+allele  in one or both parents  give frequencies of prototrophic recombinantr  that mearound  I5 times lower  than  those  in crosses  homozygous  for the recessive eallele. The recombination-3 gene  doer not  controlrecombination a+  the hirtidine-1  locus which is linked to aminotion on chromosome V or at  the hirtidine-3 locus on chromorome  I(Jho 1967  Genetics 57:365),  indicating that  its effect is  locus specific. Since y&is linked to mating type in linkage group  I,its effect on the hirtidine-5  gene  in linkage qoup  IV could be easily tested.The tests  measured recombination between the his-5 alleles  K553 and  K512. The K553 II;  y&stock  isolated  from the wildtype Em 0;  ret-3 was  crossed to each of five K512  A stocks isolated from a cross  of K512 a of unknown ret-3 constitution with thewild  type Exrec-3+. Frquencier  of prototrop&  recombinanti wiring in the progenyof  these five crosses ranged from 7.4 to- -I I.  7 per 10s  oscotporer. Since ret-3 is only 12  mop units from mating  type, the probability that ot Iart  one of the crosses  beonthe dominant ret-3+  allele  is Oxor  unity if the K512o  stock  is &. Five isolates of K512  of mating type A were isolatedfrom a cross  of o with CI  rec.3 stock cot-l (ClO2); ai  (47305), iralote no. 3675 supplied by D. G. Catcheridy  Each of- -there five isolates  wascrossed to the K553 a;  ret-3 rtockynd  the frequency of hirtidine  prototrophr in the progeny was  determined.Frequencies ranged from 8.4 to 12.6 per 105  arcospores. The probability that (I+  least  one of the five crosses wets  homorygour forrec.3 is 0.999. It  may be confidently orrumed  therefore that recombination-3 differences do not control recombination behveenwand  K512 , or if they do then the effect is only very slight.Since ret-3 controls  recombination frequency between ali pairs  of amination alleles  tested, the absence  of any  detectablecontrol of recombination between the his-5 ouxotrophr  K553 and  K512 odds considerable weight to the rupparition that controlby ret-3 is  locus specific.-Further tests  will be needed to determine whether ret-3 controlr  recombination a+  loci other thanomiXn.  - - - Deportment of Genetics, University of Leeds, Leeds 2, England.Kapoor, M. and  D. Bray. Cotobolite During on experiment deigned  to study the effect of growth conditions on theeffects on some  enzymes of Neurosporaactivity and  synthesis of glutamine rynthetore, several interesting observotionr  weremade. Neurorporo crass0  pe (Y 8743m) (FGSC  W37)  was  used  0s  D source  of enzymesin this  study. All c=sWere  prepared in Vogel’s minimal medium with sucroseor glucase  as a carbon  source,  and mycelial powders were  obtained OI described in the communication  on phaphofructokinore  inthis  iswe  of the Neurospora Newsletter. The cultures were grown  for 30 hours  at 28OC. There  WCS  no outolyrir  in culturer withlow  concentration of sucrose. At the end of 30 hours not all of the sugar in the medium was exhausted, o very small amount re-maining. Crude extracts were prepared by extracting ot 3°C lyophilired  mycelium  powder in 0.05M  phaphote  buffer (5 x 10m4M in EDTA and  IO-4 M in O-mercoptoethonol  ) pH  7.5, for 30 minutes, straining the mixture through four loyerr  of cheeseclothand  centrifuging  the supernotant  a+  27,000 x 9 for 15  minutes. The residue  was  discorded and  the rupernatant was  used  withoutfurther purif&&n.Table  I. Effect of sucrose on some  enzymes of Neurospora.Sucrore Specific activity ( OD/mg  protein)c o n c e n t r a t i o n  G D H - D  G D H  - T  GluN-Sore  P KO,l% 0.75 0.05 0.0050.75 0.05 0.0000 .5% 0.56 0.37 0. 180.53 0.38 0. I7I 0 % 0.45 0.70 0.320.43 0.70 0.321.5% 0.23 0.81 0.410.24 0.79 0.412.0% 0. I5 0.86 0.42c.  1 5 0.86 0.422 .5% 0.12 0.98 0.450. I9 0.98 0.440.120. I21.301.402.002.202 . 12. II .752.00Activities of glutomine rynthetare (GluN-S’ore),  NAD-specificglutamate dehydrogenose  (GDH-D), NADP-specific glutamate de-hydrogenare  (GDH -T) and pyruvote  kinore  (PK ) were determinedin extractr  of mycelia  obtained from cultures  ~rwn  in different con-centrotiom of wcrcse.  Arrays of the activities of the two  glutamatedehydrogenarer  were performed with a Gilford  model 2000 recordingrpectrophotometer by following the initial decrease in OD at  340 mpaccompanying the reductive ominotion of o-ketoglutarate  in the pre-sence of ommonio  and  reduced NAD or reduced NADP (K apoor  andSmith 1968 Can. J.  Microbial.  14:609).  GluN-S’are  was  arrayedby measuring the formotion of ?-glutomyl  hydroxomate  from L-glut-amote  and  hydroxylomine in the presence of ATP (K apoor  and Bray1968 Biochemistry 7:3583).  PK wcls  measured  by following the de-creore  in OD Al+  340 mp in the following reaction mixture at  25’C:Tris-HCI,  pH  8.0, 100  pm&r; M&12  IO pmoler;  ADP I pmole;reduced NAD 0.14pmole;  PEP 0.6pmole;  LDH (Sigma) IOOpg  andenzyme preporation in 0 total volwne  of 3 ml.2.59 Table 1 shows the specific activities of there enrymer in crude2.30 extrach of Neurcsporo  mycelium drown  ot concentrations varying from0. 1% to 2.5%. Glutamine  rynthetare is not repressed by wcrcse  andneither is pyruvote  kinare;  both there enzymer  rhw on increase in specific activity in the presence of wcrose  up to 1.5% but nofurther increow was  noted at 2% and  2.5% sucrose. A study of the response  of the Iwo  GDH’r  towards  wcrose  in the growth mediumrevealed  CI  dromotic  feature of regulation of GDH-T and  GDH-D. Whereor  GDH-D is subject to cotobolite  repression by sucroseand glucose, GDH-T is  induced under the same  conditions, thus demonrtroting  a reciprocal relationship between these h v o  enzymes.It  is already known that  in the presence of glutamate or ammonia in the medium GDH-D is induced with a simultoneovs  repressionof GDH-T (Sanwal  and  Loto  1962 Arch. Biochem.  Biophys.  98:420).  I t  h a s b een  suggested that GDH-D is primarily a catabolicenzyme and that GDH-T serves on anabolic function in the cell. Our studier are in agreement with this suggestion  in so  for 01  itis GDH-D alone that  is rubiect to catobolite  repression and that  GDH-T is induced under the some  conditions. - - - Deportmentof Biology, University of Calgary, Calgary, Alberta, Conodo.Van Winkle, W. B.  Preliminary obrervotionr The growth and gross morphological  features of slime have been presented byof the ultrastru~twe  of the slime  mutant.Emerson (1963 Genetics 34: 162). The heterocoryon  (fr;rg;org-I,  CT, our,  or-l )M I ) (FGSCff327),  as  well 01  th;r&thodrfor  rusta~nnng+ (*, nit-I,  lp ;slime, wexnd  y ruppxd  by V. W. Woodward.  Growth on ogor  facilitated theisolation of the hyphlets. H yphleh 24-48 hours old were  fixed in 2.5% glutoroldehyde  and  portfixed in 0~04,  both buffered with0. 16MSorensen’s  buffer. Following staining in aqueous  uronyl acetate and ethanol dehydmtion,  specimens were embedded inAraldite  for electron microscopy.Electron microscopic examination reveals that  the majority of hyphleh from spheroplartr  are devoid of cell walls. The absenceof cell wollr  no doubt clllws  for good fixation by the glutoroldehyde. Cells  ore  found to be multinucleote  with obvious connec-tions between the nuclear  envelope and  the rough endoplosmic  reticulum. Prominent granular nucleoli are  present, usually oneper nucleus.  To dote, only rough endoplormic reticulum has  been &served  in slime. Occorionolly,  the endoplarmic reticulum isfound in large lomellor  orrangementr  of 3-5 layers  of membranous  ~twctures.  Oblique or glancing sections of endoplormic retic-ulum show  numerous  polyribmomer associated with the membranes.Mitochondrio of the typical  elongate form ore very common; however, some  do exist in doughnut-like configurations. Thecristoe  of the mitochondria  terminate in square  ends or in bulbor  shaper. Serial sections have revealed that what appear  to beseveral mitochondria in single  sections are actually different lobes of single large mitochondrio. In younger hyphletr myelinwhorls involving both the inner and  outer  mitochondrial  membranes have been observed. Some mitochondrio are  seen  to hove“buds”  of the outer membrane extending into the surrounding cytoplasm.An outstanding feature of slime hyphlets is the abundance  of dense  granular  organeller, morphologicolly identical to micro-bodies  (peroxiromer  ). These organeller  range  in diameter  from 0.13~  to 0.18p and,  in many  cases,  are dumbbell-shaped  andassociated with rough endoplormic reticulum sections. Ultrostructurol demonrtrotion  of the presence of various  enzymes in per-oxisomes,  mitochondrio and  endoplarmic reticulum is currently being attempted. (Robert Welch Foundation Grant F-060. )_ _ _ Deportment of Zoology, University of Texas  at  Austin, Austin, Texas  78712.Bokerrpigel,  A. Migrating and dividing Recent HCI-Giemro and Azure A-SO2 stained  preporations  hove provided newnuclei in somatic  cells of Neurosporo.observations on dividing nuclei in somatic cells of Neurorporo craw:I) In contrast  to the report by Wilson  (1966 Neurosporo  Newrl.  lO:6),  divisiondoer not always  occur perpendicular to the longitudinal axis of the cell. In fact,nuclei in sevewl  stager of division can  be found lying parallel to the longitudinal axis  of germinating conidia and hyphol  cells.2) During  the final reparation of sister  nuclei (telophare)  two logging chromosomes con  be observed. One end of each ofthere chromosomes  is  attached  to the nucleus. Their free ends are swollen or bulbous.3) During early nuclear division the chromosomes appear to be looped, hooked or curved ot one  end. A ring-rhoped chromo-cane  her  also  been observed at  this stage. The free ends of at leort three chromosomes in metaphare  complexes were observed tohave swollen  ends.4) A migrating nucleus in CI  hypho  con be oval  in shape. A long slender  intronuclear  strand  can  be observed ottached  to thenuclear  membrane. This strand  may be “‘?-shaped  and extends away  from the reptol pore through which the nucleus  migrated,terminating in a smell,  densely rtoined  body  which is assumed  to be the centriole. Such a strand  may  be composed of two par-tionr, one of which is twice the length of the other. The regions  to which there strands  are attached  oppeor  to be composed oftiny granules situated  opposite each  other on the nuclear  membrane. In contrwt  to some previous reports, none of there strandswere  observed to extend in opposite directions.5) Although chromcsomol  complexes and  individual chromosomes were observed, no classical alignment of these chromosomeson a metophore  plate war noted. Wilson reported that  a spindle occur  only between separating chromatidr. Present okerva-tions  suggest  that  D  “spindle approtur”, similar to that described by the writer in Trichophyton mentogrophyter  (Robinow andBokerrpigel  1965 p.  119-161. In Aintworth and  Surrman  (eds), The Fungi, Vol. I.  Academic Press, New York ), my operatein dividing, somatic  nuclei of r crossa.  - - - Department of Clinical Microbiology, Victoria Horpitol,  London, Ontario,Canada.Morgan, D. H. Arginarelers  mutants The selection procedure for orginare  mutants  described by R. H. Dovir,o f  Neurorporcl(1968 Report of the Fourth Neurorporo Information Conference, NevrorporaNewrl.  13: p.  13) was arrived  ot independently by the author while workingin N. H. Horowitz’s laboratory ot Posodena.  Gvr  procedures differed in onlytwo  respects: I)  I used Vogel’s “N”, with wmnonio  OI nitrogen scurce,  for the filtration and  plating medium whereas DavCanticipated complications involving orginine uptake in the presence of ammonia and  therefore used  o nitrate minimal.

Catabolite effects on enzymes

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Catabolite effects on enzymes

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