< 



Z 4 6 8 10 



DAYS AFTER TREATMENT 

 RNA and DNA content of buds of potato 

 tubers at varying times after 3-day pretreatment with 

 ethylene chlorohydrin. 



Fig. 13. 



(Fig. lA, Tuan and Bonner, Plant Physiol. 39, 768, 1964; 

 reproduced with the permission of the American Society 

 of Plant Physiologists.) 



E 

 CM 



d 



< 



IT 

 O 



a. 

 a: 

 o 



CJ 



z 

 a. 

 < 



4. 



10 20 30 



|ig DNA per 0.25 ml 



Fig. 14. 



Template activity of rat Uver chromatin Isolated 4 hours 

 after treatment with hydrocortisone(-o-)or saline (-a-). 



histories to control was as follows. It has been 

 reported recently in the literature that cells 

 can be treated with hormones in such a way as 

 to give a histone-hormone linkage, and it was 

 implied that hormones might be pulling the his- 

 tones off DNA. We had an excellent system with 

 which to examine this hypothesis since we were 

 able to study large amounts of target organ 

 tissue. 



We were anxious to see if endometrial tissue 

 incubated in vitro followed some of the rules 

 that one would expect from the in vivo endome- 

 trial material. Figure 15 gives an account of the 

 uptake of hormones into the endometrial cell. 

 There appears to be some degree of additional 

 concentration of estradiol and progesterone. 

 Progesterone is also a hormone which has the 

 endometrium as a target tissue during preg- 

 nancy, and so it is not surprising that it is also 

 concentrated into the tissue. Incorporation of 

 hydrocortisone which, of course, has liver as its 

 target organ was low. In Fig. 16 you see the 

 uptake into the cytoplasmic fraction. It follows 

 the overall pattern of the previous figure. How- 

 ever, now when we looked at the lysed nuclei 

 [which I will refer to as crude chromatin (bottom, 

 Fig. 16)] we began to see a very dramatic dif- 

 ference. Again, I stress that as yet lam discus- 

 sing hormone uptake and not binding. There are 

 large amounts present of the hormones for which 



TABLE IV 



Effectiveness of Chromatin of Dormant and of 

 Non-dormant Potato Buds in the Support of DNA- 

 dependent RNA Synthesis by Exogenous RNA Polymerase 



For composition of reaction mixture see Materials and 

 Methods. 



50 Mg of DNA 

 supplied to system as : 



RNA synthesized 



/iyumole AMP incorp 



per 10 min 



Potato DNA (deproteinized) 

 Chromatin of potato tuber 

 Chromatin of dormant buds 

 Chromatin of buds from tubers at 



end of 3-day treatment with 



ethylene chlorohydrin 

 Chromatin of buds from tubers 



10 days after 3-day treatment 



with ethylene chlorohydrin 



3370* 

 

 122 



1412 

 1538 



* Incorporation due to polymerase alone (150 /u/imole) 

 subtracted. 



(Table I. Tuan and Bonner. Plant Physiol. 39, 768, 1964; 

 reproduced with permission of the American Society of 

 Plant Physiologists.) 



this is the target tissue, and small amounts of 

 the other steroids. I should add that the specific 

 activity of testosteroneandestradiol were within 

 2% of each other. Hydrocortisone was somewhat 



141 



